Vivian, this is not a place to advertise but rather to share knowledge and hash things our. Should want to continue to participate, please do not post things about your company's services or products. Thank you.
Posts in Antibody Discussion Forum
Exosome antibodies are antibodies developed against exosomal surface markers. They have been widely used as exosome research tools and for clinical applications such as disease prognosis and diagnosis. To help promote the exosomal studies and their clinical use, Creative Biolabs provides high-quality antibody development services using different technologies, including phage display antibody screening and B cell sorting, which can help the discovery of antibodies that recognize exosome-associated antigens with sufficient sensitivity and specificity. https://www.creative-biolabs.com/exosome/exosome-antibody-development-services.htm
A trip to Tibet is different from a trip to any other place on earth. Tibet is a land of four distinct seasons, with a monsoon in summer, which brings much less rainfall than other monsoon countries. Its landscapes range from mountains to valleys. Temperatures can change drastically, with pleasantly warm temperatures during the day dropping to freezing at night, even in summer. Mastery the basics to get <a href="https://www.greattibettour.com/top-7-tibet-tours">the Best Tibet tours experience</a>. Unique Climate and Weather One of the most unique things about Tibet is the climate. Tibet has four seasons: spring, summer, autumn and winter. Summers can be hot in some areas and cooler in others, while winters are generally cold. Spring and autumn are the best time for travel in Tibet; the weather is relatively mild, which is very suitable for tourism and trekking, which are the top activities in Tibet. Unique Religion Tibet has been a major Buddhist region for over 1300 years. In Tibet, Buddhism is not just a religion. It is a complete way of life. Tibetan culture is a culture that fully integrates Buddhism into its fabric, and Tibetans bring Buddhism into their lives every day, and everything they do. Outside Lhasa One of the most important things about Tibet is the altitude. The average altitude of the Tibetan Plateau is over 4,800 meters above sea level, and the thin air can cause altitude sickness, although the body can usually adjust to the reduced oxygen in a day or two to compensate. From Lhasa, the capital of Tibet, to the base camp of Mount Everest, Mount Kailash, and Lake Namtso, the altitude is often worrying. In Tibet, you need to understand that altitude sickness can happen, but you will overcome it and get used to it. It can take a whole day to get from one town to another. There are many customs in Tibet that are part of the Buddhist belief, so you need to watch your words and actions so as not to offend.
Thanks Frances. I appreciate your input. Kind regards. Shadi Shadi Bootorabi Monoclonal Antibody Senior Biological Scientist UF ICBR 2033 Mowry Road, Gainesville, FL 32610 Biotech.ufl.edu | Tel: 352.273.8041| Fax: 352.273.8070
-----Original Message-----
From: Antibody Discussion Forum <email obscured>> On Behalf Of
<email obscured>
Sent: Thursday, February 10, 2022 11:12 AM
To: <email obscured>
Cc: Angel Sampson <email obscured>>
Subject: Re: [ABRF Ab Forum] recommendation for electrofusion instrument
[External Email]
We use the Nepa Gene Super Electro Cell Fusion Generator only because the tech
support was better then BTX ... but both units work well. My staff tried them
out. There are no consumables with Nepa Gene's, so if BTXs fusion chambers are
disposable, it will be more expensive in the end as well. Pretty sure we buy
BTX electro fusion buffer ... but I would need to check that.
Frances
Office: 646-888-2354
Mobile: 516-967-0656
Questions: <email obscured>
When publishing your work, please remember to:
Acknowledge support from the Cancer Center Support Grant (2 P30
CA008748-53)
Include the Coreβs RRID # (CR 017691)
-----Original Message-----
From: Antibody Discussion Forum <email obscured>> On Behalf Of
Bootorabi,Shadi
Sent: Thursday, February 10, 2022 10:02 AM
To: <email obscured>
Cc: Angel Sampson <email obscured>>
Subject: [EXTERNAL] [ABRF Ab Forum] recommendation for electrofusion instrument
β 1 file link β
Good morning everybody
We are shopping around for a good electrofusion system . What is the best
instrument you recommend?
I appreciate your feedback. Best regards. Shadi
Shadi Bootorabi
Monoclonal Antibody Senior Biological Scientist
UF ICBR
2033 Mowry Road, Gainesville, FL 32610
Biotech.ufl.edu<https://www.biotech.ufl.edu/> | Tel: 352.273.8041| Fax:
352.273.8070
[cid:image001.png@01D81E65.325103D0]
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We use the Nepa Gene Super Electro Cell Fusion Generator only because the tech
support was better then BTX ... but both units work well. My staff tried them
out. There are no consumables with Nepa Gene's, so if BTXs fusion chambers are
disposable, it will be more expensive in the end as well. Pretty sure we buy
BTX electro fusion buffer ... but I would need to check that.
Frances
Office: 646-888-2354
Mobile: 516-967-0656
Questions: <email obscured>
When publishing your work, please remember to:
Acknowledge support from the Cancer Center Support Grant (2 P30
CA008748-53)
Include the Coreβs RRID # (CR 017691)
-----Original Message----- From: Antibody Discussion Forum <email obscured>> On Behalf Of Bootorabi,Shadi Sent: Thursday, February 10, 2022 10:02 AM To: <email obscured> Cc: Angel Sampson <email obscured>> Subject: [EXTERNAL] [ABRF Ab Forum] recommendation for electrofusion instrument β 1 file link β Good morning everybody We are shopping around for a good electrofusion system . What is the best instrument you recommend? I appreciate your feedback. Best regards. Shadi Shadi Bootorabi Monoclonal Antibody Senior Biological Scientist UF ICBR 2033 Mowry Road, Gainesville, FL 32610 Biotech.ufl.edu<https://www.biotech.ufl.edu/> | Tel: 352.273.8041| Fax: 352.273.8070 [cid:image001.png@01D81E65.325103D0] β 1 file β β‘ image001.png (9kb) http://list.abrf.org/r/file/lygPs6UdEfrqCBGdI1YgIUOXnr6-2mY-2TvjHi8/ ββ View topic http://list.abrf.org/r/topic/3iofrPik2HhOy2SrWwdV10 Leave group <email obscured>?Subject=Unsubscribe *** Only open attachments or links from trusted senders. Report phishing to <email obscured> *** ===================================================================== Please note that this e-mail and any files transmitted from Memorial Sloan Kettering Cancer Center may be privileged, confidential, and protected from disclosure under applicable law. If the reader of this message is not the intended recipient, or an employee or agent responsible for delivering this message to the intended recipient, you are hereby notified that any reading, dissemination, distribution, copying, or other use of this communication or any of its attachments is strictly prohibited. If you have received this communication in error, please notify the sender immediately by replying to this message and deleting this message, any attachments, and all copies and backups from your computer.
Good morning everybody We are shopping around for a good electrofusion system . What is the best instrument you recommend? I appreciate your feedback. Best regards. Shadi Shadi Bootorabi Monoclonal Antibody Senior Biological Scientist UF ICBR 2033 Mowry Road, Gainesville, FL 32610 Biotech.ufl.edu<https://www.biotech.ufl.edu/> | Tel: 352.273.8041| Fax: 352.273.8070
[cid:image001.png@01D81E65.325103D0]
Posting here in case one of us has a colleague with antibody drug discovery experience and would consider working in MSKCC's Office of Technology Development ... please share this with them: Senior Lead, Antibodies & Biologics Drug Development We Are: β’ The Office of Technology Development's (OTD) mission is to ensure that Memorial Sloan Kettering's (MSK) inventions have the greatest chance of being developed and commercialized - so that these inventions both benefit patients and return the most value to MSK. β’ The Senior Lead, Antibodies & Biologics Drug Development plays a crucial role in fulfilling this mission. You Will: β’ Working with the Licensing Manager, the Senior Lead will lead generation and execution of drug development plans and strategies and will be a resource for advising on leading drug development technologies and service providers. β’ The Senior Lead will further serve as point-person in relationships with internal and external collaborators (including consultants, CROs, and core facilities) that are collaborating with MSK on drug development projects. β’ The Senior Lead will act as liaison between investigators and different groups at MSK to manage such drug development projects and ensure their timely and efficient progression to clinic. You Need: β’ Master's degree in Life Sciences (PhD a strong plus) β’ At least 4 years experience (2 years with PhD) in therapeutic drug development and project management, preferably in an industry or health care environment β’ Demonstrated analytical skills β’ Strong communication skills, written and oral. β’ Strength in collaboration: able to work effectively with both industrial and academic colleagues. β’ Ability to simultaneously handle multiple priorities & projects to successful completion
Happy Holidays all. I use our in-house peptide facility here. Sorry! I wish all of you and yours an amazing 2022. Best,
Scott -----Original Message----- From: Antibody Discussion Forum <email obscured>> On Behalf Of Frances Weis-Garcia Sent: Wednesday, December 22, 2021 11:42 AM To: <email obscured> Subject: [ABRF Ab Forum] Recommendations - Peptide Synthesis Caution: External Sender. Do not open unless you know the content is safe. Any recommendations on cores or companies that synthesize peptides? Who do you use / trust? Hope you all have Blessed and Joyous Holiday Season as well as a Brighter 2022! Be well everyone. ββ View topic https://nam11.safelinks.protection.outlook.com/?url=http%3A%2F%2Flist.abrf.org%2Fr%2Ftopic%2F7mbvcLh9kDdNVfTK2ngS6q&data=04%7C01%7Cscott.brown%40stjude.org%7C2a5fdc83cb9d43946f8c08d9c5725df1%7C22340fa892264871b677d3b3e377af72%7C0%7C0%7C637757917694029232%7CUnknown%7CTWFpbGZsb3d8eyJWIjoiMC4wLjAwMDAiLCJQIjoiV2luMzIiLCJBTiI6Ik1haWwiLCJXVCI6Mn0%3D%7C3000&sdata=woB3d1TdtkYhIO%2FEYJ684tVYxOg3u1u%2BKqnFVJK1KcE%3D&reserved=0 Leave group <email obscured>?Subject=Unsubscribe ________________________________ Email Disclaimer: www.stjude.org/emaildisclaimer Consultation Disclaimer: www.stjude.org/consultationdisclaimer
Any recommendations on cores or companies that synthesize peptides? Who do you use / trust? Hope you all have Blessed and Joyous Holiday Season as well as a Brighter 2022! Be well everyone.
vivian.creativebios
Job Posting - Principal Scientist, Modelorg Antibody Discovery and Engineering in New York
Posted at
Dear All, Creative Biolabs is looking for a motivated and experienced Principal Scientist specialized in Model Organism Antibody Discovery and Engineering. This position will lead antibody discovery and engineering efforts across various programs at Creative Biolabs, with an initial focus on model organisms. The candidate is expected to provide well-researched insights with technical knowledge and analytical skills that will influence Creative Biolabs' technology development. The company website: https://modelorg-ab.creative-biolabs.com/ The position introduction page: https://modelorg-ab.creative-biolabs.com/careers.htm
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Sabin, Unfortunately we do not do single B cell cloning. Hopefully someone on this discussion forum does ... but in the mean time let's think this through together. So when you say you are having problems expressing functional antibodies from the sorted B cells, what exactly do you mean? Are you able to clone out and express antibody but none are specific for the target? If so, have you done a control where you take half the spleen, fuse it and find the antigen specific MAb in the traditionally way and in parallel do you single B cell cloning approach. If you do not get them with either approach, you know it was the source of B cells. If you get them with the traditional hybridomas, it is the single B cell cloning protocol.
Frances
____________________
ο»ΏOn 11/1/21, 1:39 PM, "Antibody Discussion Forum on behalf of Sabin.Antony"
<email obscured> on behalf of <email obscured>> wrote:
Hi I had some issues expressing functional antibodies from target sorted
B cells. We expressed antibody sequences obtained by both single cell
sequencing and bulk B cell sequencing. I was expecting ~80% successful rate but
not much luck so far. Any suggestion you can offer will be really helpful.
Thanks
ββ
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Probably you can reach some professional service providers. Do not mean to spam, for single-cell sequencing, you may want to try: https://www.creative-biolabs.com/suprecision/suprecision-platform.htm
Hi I had some issues expressing functional antibodies from target sorted B cells. We expressed antibody sequences obtained by both single cell sequencing and bulk B cell sequencing. I was expecting ~80% successful rate but not much luck so far. Any suggestion you can offer will be really helpful. Thanks
Anyone have an idea on how to monitor for the success of an IV injection or better yet IV boost? I ask because I am thinking that the variability in the # of hits and IgGs in our fusions of well immunized mice (high titers) correlates with how well the IV boost is done. 2 weeks ago we boosted and fused 2 mice (2 different immunogens ... both ion channels ... both made by the same lab) that were hyperimmunized (OD at 1:100,000 was > 0.5). One got a boat load of hits, while we only got a few for the other ... some of this correlates with the number of IgGs ... but not all of it. They really should have both been win falls. I am not surprised because this happens all the time ... but it should "NOT" ... and I am wondering if the IV boost has something to do with it. I know you can see the blood go away in the vein when it gets into the vein, but I am wondering if there is a better way to access how good the IV boost was received ... like seeing a tracer dye in the translucent ears ... being a little silly now I know.
Thoughts?
Thanks ... I'll reach out to him.
ο»ΏOn 8/10/21, 1:43 PM, "Antibody Discussion Forum on behalf of David Blum"
<email obscured> on behalf of <email obscured>> wrote:
There was a seminar from the MADSSCi group today and one of the speakers
was from Cytiva and spoke on magbeads for DNA. He said he could talk to
anyone that had questions about implementing magnetic beads into their
workflow for antibody purification. His contact info and Cytiva's product
is below.
<email obscured>
https://www.cytivalifesciences.com/en/us/shop/protein-analysis/protein-sample-preparation/protein-enrichment/protein-g-mag-sepharose-p-01111
On Tue, Aug 10, 2021 at 10:33 AM Weis-Garcia, Frances/Sloan Kettering
Institute <email obscured>> wrote:
> Thanks everyone!
>
> Frances
>
> ZRC 1563
> Office 646-888-2354
> Mobile 516-967-0656
>
>
> On 8/10/21, 10:22 AM, "Antibody Discussion Forum on behalf of
> Bover,Laura" <email obscured> on behalf of <email obscured>>
> wrote:
>
> We collect 12 aliquots of 1 ml each and then we measure in nanodrop
> the OD. Usually the peak appears in the 4th-5th elution sample. We pool
as
> need it. Filter and eventually concentrate.
>
> Thanks!
>
> On 8/10/21, 9:14 AM, "Antibody Discussion Forum on behalf of David
> Blum" <email obscured> on behalf of <email obscured>> wrote:
>
> WARNING: This email originated from outside of MD Anderson.
Please
> validate the sender's email address before clicking on links or
attachments
> as they may not be safe.
>
> Frances,
>
> If you don't want to use an FPLC you can use a simple syringe
with
> the
> pre-packed protein a or g columns and a luer adapter which may be
> included
> in the box with the column. Since this is a bind an elute
process
> it
> should be fine to do it this way, but you would need to determine
> the
> optimal elution volume and range since you won't have a UV
> detector.
>
> On Mon, Aug 9, 2021 at 10:54 AM Weis-Garcia, Frances/Sloan
> Kettering
> Institute <email obscured>> wrote:
>
> > Thanks David.
> >
> > Frances
> >
> > ZRC 1563
> > Office 646-888-2354
> > Mobile 516-967-0656
> >
> >
> > On 8/8/21, 1:28 PM, "Antibody Discussion Forum on behalf of
> David Blum" <
> > <email obscured> on behalf of <email obscured>> wrote:
> >
> > Frances: I understand. This might not be the best option,
> but it
> > should
> > be good for processing multiple samples quickly without
> using the FPLC.
> > You could also batch bind with the resin you use now and
> spin down at a
> > speed that doesn't deform the beads. Good luck!
> >
> > David
> >
> > On Sat, Aug 7, 2021 at 1:30 PM Weis-Garcia, Frances/Sloan
> Kettering
> > Institute <email obscured>> wrote:
> >
> > > Thanks David. The binding capacity is pretty low
relative
> to
> > standard
> > > resins and the beads are much more expensive.
> > >
> > > > 0.6 mg rabbit IgG / ml Pierceβ’ Protein G Magnetic
> Beads
> > >
> > > while
> > >
> > > > 20.0 mg human IgG / ml Cytiva Protein G Sepharoseβ’ 4
> Fast Flow
> > Media
> > > (formerly GE LifeSciences / Formerly Pharmacia ... boy am
> I old)
> > >
> > > I know I want to use the FastFlow resin (or something
> similar) ...
> > right
> > > now I am more looking for a process that does not involve
> an FPLC if
> > I can
> > > help it. I have one but the person who has used it is
> retireing and
> > this
> > > is the only thing we will use it for, so do I need it is
> the
> > question.
> > >
> > > A lab (Ivo this was Pop Council) used to grow up a 100s
of
> mls of
> > each
> > > hybridoma then line the bottles of conditioned media up
on
> a shelf,
> > > allowing the media to flow through a tube into a column
> below it and
> > pass
> > > over the a prot G resin over night by gravity. Some how
> they set it
> > up so
> > > the columns DID NOT DRY OUT. The next morning they would
> wash the
> > columns
> > > by gravitty again in series and then elute them one at a
> time.
> > Sounds like
> > > a great way to do a 5 to 20 at once, and I am sure I
could
> figure
> > this out,
> > > but before I spend time on this, was wondering if anyone
> else already
> > > tackled this or had a better mouse trap.
> > >
> > > Frances
> > >
> > > ZRC 1563
> > > Office 646-888-2354
> > > Mobile 516-967-0656
> > >
> > >
> ____________________________________________________________
> > >
> > > On 8/6/21, 7:41 PM, "Antibody Discussion Forum on behalf
> of David
> > Blum" <
> > > <email obscured> on behalf of <email obscured>>
> wrote:
> > >
> > > Have you tried mag beads? We use them for histagged
> proteins at
> > small
> > > scale but there are protein coupled beads too.
> > >
> > >
>
https://urldefense.com/v3/__https://www.thermofisher.com/order/catalog/product/88848__;!!PfbeBCCAmug!3WJ0-E0eUzj5cM-MvKLQ5OyZru8t7KhqsTPZP-Gh4ptWUlPQugH9vEtHLaTYxzoe$
> > >
> > >
> > >
> > > On Fri, Aug 6, 2021 at 6:47 PM Weis-Garcia,
> Frances/Sloan
> > Kettering
> > > Institute <email obscured>> wrote:
> > >
> > > > Happy Friday.
> > > >
> > > > Do any of you produce and purify small amounts of
> the new MAbs
> > you
> > > make
> > > > for further characterization prior to committing to
> a few or
> > one
> > > winner?
> > > > If so, how do you do this. Currently we make 350
ml
> exhausted
> > media
> > > > (hybridoma adapted to 1% FBS prescreened for
> ultra-low levels
> > of
> > > bovine IgG
> > > > + 1 ng/ml IL6) and then purify it on a Prot G
column
> using and
> > > FPLC. We
> > > > get 1 β 8 mg, but I would like to scale this back
> and simply
> > it if
> > > possible
> > > > because we usually do not need that much.
> > > >
> > > > Thanks.
> > > >
> > > > Frances
> > > >
> > > >
> > > > Frances Weis-Garcia, Ph.D.
> > > >
> > > > Head
> > > > Bi-Institutional Antibody and Bioresource Core
> Facility
> > > > (RRID: CR 017691)
> > > >
> > > > Associate Laboratory Member
> > > > Immunology Program
> > > >
> > > > Memorial Sloan Kettering Cancer Center
> > > > 1275 York Avenue, New York, NY 10065
> > > > <
> > >
> >
>
https://urldefense.com/v3/__https://www.google.com/maps/search/1275*York*Avenue,*New*York,*NY*10065?entry=gmail&source=g__;KysrKysr!!PfbeBCCAmug!3WJ0-E0eUzj5cM-MvKLQ5OyZru8t7KhqsTPZP-Gh4ptWUlPQugH9vEtHLcA59hJ3$
> > > >
> > > > T 646.888.2354 C 516.967.0656
> > > >
> > > > ABCF-MSKCC website<
> > > >
> > >
> >
>
https://urldefense.com/v3/__https://www.mskcc.org/research-advantage/core-facilities/monoclonal-antibody-core-facility__;!!PfbeBCCAmug!3WJ0-E0eUzj5cM-MvKLQ5OyZru8t7KhqsTPZP-Gh4ptWUlPQugH9vEtHLfRBPtf4$
> > > > >
> > > >
> > > > Adjunct Faculty
> > > > Center for Clinical and Translational Science
> > > >
> > > > The Rockefeller University
> > > > 1230 York Avenue, New York, NY 10065
> > > > <
> > >
> >
>
https://urldefense.com/v3/__https://www.google.com/maps/search/1230*York*Avenue,*New*York,*NY*10065?entry=gmail&source=g__;KysrKysr!!PfbeBCCAmug!3WJ0-E0eUzj5cM-MvKLQ5OyZru8t7KhqsTPZP-Gh4ptWUlPQugH9vEtHLZ2GrQuq$
> > > >
> > > > T 212.327.7030
> > > >
> > > > ABCF-RU website<
>
https://urldefense.com/v3/__https://www.rockefeller.edu/monoclonal/__;!!PfbeBCCAmug!3WJ0-E0eUzj5cM-MvKLQ5OyZru8t7KhqsTPZP-Gh4ptWUlPQugH9vEtHLbgJ5Ak4$
> >
> > > >
> > > >
> > > > Please note that this e-mail and any files
> transmitted with it
> > may be
> > > > privileged, confidential, and protected from
> disclosure under
> > > applicable
> > > > law. If the reader of this message is not the
> intended
> > recipient, or
> > > an
> > > > employee or agent responsible for delivering this
> message to
> > the
> > > intended
> > > > recipient, you are hereby notified that any
reading,
> > dissemination,
> > > > distribution, copying, or other use of this
> communication or
> > any of
> > > its
> > > > attachments is strictly prohibited. If you have
> received this
> > > > communication in error, please notify the sender
> immediately by
> > > replying to
> > > > this message and deleting this message, any
> attachments, and
> > all
> > > copies and
> > > > backups from your computer.
> > > >
> > > >
> > > >
> >
> =====================================================================
> > > >
> > > > Please note that this e-mail and any files
> transmitted
> > from
> > > > Memorial Sloan Kettering Cancer Center may be
> privileged,
> > > > confidential,
> > > > and protected from disclosure under applicable
> law. If the
> > > reader of
> > > > this message is not the intended recipient, or
> an
> > employee or
> > > agent
> > > > responsible for delivering this message to the
> intended
> > > recipient,
> > > > you are hereby notified that any reading,
> dissemination,
> > > distribution,
> > > > copying, or other use of this communication or
> any of its
> > > attachments
> > > > is strictly prohibited. If you have received
> this
> > > communication in
> > > > error, please notify the sender immediately by
> replying
> > to this
> > > > message
> > > > and deleting this message, any attachments,
and
> all
> > copies and
> > > backups
> > > > from your computer.
> > > >
> > > > ββ
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There was a seminar from the MADSSCi group today and one of the speakers was from Cytiva and spoke on magbeads for DNA. He said he could talk to anyone that had questions about implementing magnetic beads into their workflow for antibody purification. His contact info and Cytiva's product is below. <email obscured> https://www.cytivalifesciences.com/en/us/shop/protein-analysis/protein-sample-preparation/protein-enrichment/protein-g-mag-sepharose-p-01111 On Tue, Aug 10, 2021 at 10:33 AM Weis-Garcia, Frances/Sloan Kettering
Institute <email obscured>> wrote: > Thanks everyone! > > Frances > > ZRC 1563 > Office 646-888-2354 > Mobile 516-967-0656 > > > ο»ΏOn 8/10/21, 10:22 AM, "Antibody Discussion Forum on behalf of > Bover,Laura" <email obscured> on behalf of <email obscured>> > wrote: > > We collect 12 aliquots of 1 ml each and then we measure in nanodrop > the OD. Usually the peak appears in the 4th-5th elution sample. We pool as > need it. Filter and eventually concentrate. > > Thanks! > > On 8/10/21, 9:14 AM, "Antibody Discussion Forum on behalf of David > Blum" <email obscured> on behalf of <email obscured>> wrote: > > WARNING: This email originated from outside of MD Anderson. Please > validate the sender's email address before clicking on links or attachments > as they may not be safe. > > Frances, > > If you don't want to use an FPLC you can use a simple syringe with > the > pre-packed protein a or g columns and a luer adapter which may be > included > in the box with the column. Since this is a bind an elute process > it > should be fine to do it this way, but you would need to determine > the > optimal elution volume and range since you won't have a UV > detector. > > On Mon, Aug 9, 2021 at 10:54 AM Weis-Garcia, Frances/Sloan > Kettering > Institute <email obscured>> wrote: > > > Thanks David. > > > > Frances > > > > ZRC 1563 > > Office 646-888-2354 > > Mobile 516-967-0656 > > > > > > On 8/8/21, 1:28 PM, "Antibody Discussion Forum on behalf of > David Blum" < > > <email obscured> on behalf of <email obscured>> wrote: > > > > Frances: I understand. This might not be the best option, > but it > > should > > be good for processing multiple samples quickly without > using the FPLC. > > You could also batch bind with the resin you use now and > spin down at a > > speed that doesn't deform the beads. Good luck! > > > > David > > > > On Sat, Aug 7, 2021 at 1:30 PM Weis-Garcia, Frances/Sloan > Kettering > > Institute <email obscured>> wrote: > > > > > Thanks David. The binding capacity is pretty low relative > to > > standard > > > resins and the beads are much more expensive. > > > > > > > 0.6 mg rabbit IgG / ml Pierceβ’ Protein G Magnetic > Beads > > > > > > while > > > > > > > 20.0 mg human IgG / ml Cytiva Protein G Sepharoseβ’ 4 > Fast Flow > > Media > > > (formerly GE LifeSciences / Formerly Pharmacia ... boy am > I old) > > > > > > I know I want to use the FastFlow resin (or something > similar) ... > > right > > > now I am more looking for a process that does not involve > an FPLC if > > I can > > > help it. I have one but the person who has used it is > retireing and > > this > > > is the only thing we will use it for, so do I need it is > the > > question. > > > > > > A lab (Ivo this was Pop Council) used to grow up a 100s of > mls of > > each > > > hybridoma then line the bottles of conditioned media up on > a shelf, > > > allowing the media to flow through a tube into a column > below it and > > pass > > > over the a prot G resin over night by gravity. Some how > they set it > > up so > > > the columns DID NOT DRY OUT. The next morning they would > wash the > > columns > > > by gravitty again in series and then elute them one at a > time. > > Sounds like > > > a great way to do a 5 to 20 at once, and I am sure I could > figure > > this out, > > > but before I spend time on this, was wondering if anyone > else already > > > tackled this or had a better mouse trap. > > > > > > Frances > > > > > > ZRC 1563 > > > Office 646-888-2354 > > > Mobile 516-967-0656 > > > > > > > ____________________________________________________________ > > > > > > On 8/6/21, 7:41 PM, "Antibody Discussion Forum on behalf > of David > > Blum" < > > > <email obscured> on behalf of <email obscured>> > wrote: > > > > > > Have you tried mag beads? We use them for histagged > proteins at > > small > > > scale but there are protein coupled beads too. > > > > > > > https://urldefense.com/v3/__https://www.thermofisher.com/order/catalog/product/88848__;!!PfbeBCCAmug!3WJ0-E0eUzj5cM-MvKLQ5OyZru8t7KhqsTPZP-Gh4ptWUlPQugH9vEtHLaTYxzoe$ > > > > > > > > > > > > On Fri, Aug 6, 2021 at 6:47 PM Weis-Garcia, > Frances/Sloan > > Kettering > > > Institute <email obscured>> wrote: > > > > > > > Happy Friday. > > > > > > > > Do any of you produce and purify small amounts of > the new MAbs > > you > > > make > > > > for further characterization prior to committing to > a few or > > one > > > winner? > > > > If so, how do you do this. Currently we make 350 ml > exhausted > > media > > > > (hybridoma adapted to 1% FBS prescreened for > ultra-low levels > > of > > > bovine IgG > > > > + 1 ng/ml IL6) and then purify it on a Prot G column > using and > > > FPLC. We > > > > get 1 β 8 mg, but I would like to scale this back > and simply > > it if > > > possible > > > > because we usually do not need that much. > > > > > > > > Thanks. > > > > > > > > Frances > > > > > > > > > > > > Frances Weis-Garcia, Ph.D. > > > > > > > > Head > > > > Bi-Institutional Antibody and Bioresource Core > Facility > > > > (RRID: CR 017691) > > > > > > > > Associate Laboratory Member > > > > Immunology Program > > > > > > > > Memorial Sloan Kettering Cancer Center > > > > 1275 York Avenue, New York, NY 10065 > > > > < > > > > > > https://urldefense.com/v3/__https://www.google.com/maps/search/1275*York*Avenue,*New*York,*NY*10065?entry=gmail&source=g__;KysrKysr!!PfbeBCCAmug!3WJ0-E0eUzj5cM-MvKLQ5OyZru8t7KhqsTPZP-Gh4ptWUlPQugH9vEtHLcA59hJ3$ > > > > > > > > T 646.888.2354 C 516.967.0656 > > > > > > > > ABCF-MSKCC website< > > > > > > > > > > https://urldefense.com/v3/__https://www.mskcc.org/research-advantage/core-facilities/monoclonal-antibody-core-facility__;!!PfbeBCCAmug!3WJ0-E0eUzj5cM-MvKLQ5OyZru8t7KhqsTPZP-Gh4ptWUlPQugH9vEtHLfRBPtf4$ > > > > > > > > > > > > > Adjunct Faculty > > > > Center for Clinical and Translational Science > > > > > > > > The Rockefeller University > > > > 1230 York Avenue, New York, NY 10065 > > > > < > > > > > > https://urldefense.com/v3/__https://www.google.com/maps/search/1230*York*Avenue,*New*York,*NY*10065?entry=gmail&source=g__;KysrKysr!!PfbeBCCAmug!3WJ0-E0eUzj5cM-MvKLQ5OyZru8t7KhqsTPZP-Gh4ptWUlPQugH9vEtHLZ2GrQuq$ > > > > > > > > T 212.327.7030 > > > > > > > > ABCF-RU website< > https://urldefense.com/v3/__https://www.rockefeller.edu/monoclonal/__;!!PfbeBCCAmug!3WJ0-E0eUzj5cM-MvKLQ5OyZru8t7KhqsTPZP-Gh4ptWUlPQugH9vEtHLbgJ5Ak4$ > > > > > > > > > > > > > > Please note that this e-mail and any files > transmitted with it > > may be > > > > privileged, confidential, and protected from > disclosure under > > > applicable > > > > law. If the reader of this message is not the > intended > > recipient, or > > > an > > > > employee or agent responsible for delivering this > message to > > the > > > intended > > > > recipient, you are hereby notified that any reading, > > dissemination, > > > > distribution, copying, or other use of this > communication or > > any of > > > its > > > > attachments is strictly prohibited. If you have > received this > > > > communication in error, please notify the sender > immediately by > > > replying to > > > > this message and deleting this message, any > attachments, and > > all > > > copies and > > > > backups from your computer. > > > > > > > > > > > > > > > ===================================================================== > > > > > > > > Please note that this e-mail and any files > transmitted > > from > > > > Memorial Sloan Kettering Cancer Center may be > privileged, > > > > confidential, > > > > and protected from disclosure under applicable > law. 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Thanks everyone!
Frances
ZRC 1563
Office 646-888-2354
Mobile 516-967-0656
ο»ΏOn 8/10/21, 10:22 AM, "Antibody Discussion Forum on behalf of Bover,Laura"
<email obscured> on behalf of <email obscured>> wrote:
We collect 12 aliquots of 1 ml each and then we measure in nanodrop the OD.
Usually the peak appears in the 4th-5th elution sample. We pool as need it.
Filter and eventually concentrate.
Thanks!
On 8/10/21, 9:14 AM, "Antibody Discussion Forum on behalf of David Blum"
<email obscured> on behalf of <email obscured>> wrote:
WARNING: This email originated from outside of MD Anderson. Please
validate the sender's email address before clicking on links or attachments as
they may not be safe.
Frances,
If you don't want to use an FPLC you can use a simple syringe with the
pre-packed protein a or g columns and a luer adapter which may be
included
in the box with the column. Since this is a bind an elute process it
should be fine to do it this way, but you would need to determine the
optimal elution volume and range since you won't have a UV detector.
On Mon, Aug 9, 2021 at 10:54 AM Weis-Garcia, Frances/Sloan Kettering
Institute <email obscured>> wrote:
> Thanks David.
>
> Frances
>
> ZRC 1563
> Office 646-888-2354
> Mobile 516-967-0656
>
>
> On 8/8/21, 1:28 PM, "Antibody Discussion Forum on behalf of David
Blum" <
> <email obscured> on behalf of <email obscured>> wrote:
>
> Frances: I understand. This might not be the best option, but
it
> should
> be good for processing multiple samples quickly without using the
FPLC.
> You could also batch bind with the resin you use now and spin
down at a
> speed that doesn't deform the beads. Good luck!
>
> David
>
> On Sat, Aug 7, 2021 at 1:30 PM Weis-Garcia, Frances/Sloan
Kettering
> Institute <email obscured>> wrote:
>
> > Thanks David. The binding capacity is pretty low relative to
> standard
> > resins and the beads are much more expensive.
> >
> > > 0.6 mg rabbit IgG / ml Pierceβ’ Protein G Magnetic Beads
> >
> > while
> >
> > > 20.0 mg human IgG / ml Cytiva Protein G Sepharoseβ’ 4 Fast
Flow
> Media
> > (formerly GE LifeSciences / Formerly Pharmacia ... boy am I
old)
> >
> > I know I want to use the FastFlow resin (or something similar)
...
> right
> > now I am more looking for a process that does not involve an
FPLC if
> I can
> > help it. I have one but the person who has used it is
retireing and
> this
> > is the only thing we will use it for, so do I need it is the
> question.
> >
> > A lab (Ivo this was Pop Council) used to grow up a 100s of mls
of
> each
> > hybridoma then line the bottles of conditioned media up on a
shelf,
> > allowing the media to flow through a tube into a column below
it and
> pass
> > over the a prot G resin over night by gravity. Some how they
set it
> up so
> > the columns DID NOT DRY OUT. The next morning they would wash
the
> columns
> > by gravitty again in series and then elute them one at a time.
> Sounds like
> > a great way to do a 5 to 20 at once, and I am sure I could
figure
> this out,
> > but before I spend time on this, was wondering if anyone else
already
> > tackled this or had a better mouse trap.
> >
> > Frances
> >
> > ZRC 1563
> > Office 646-888-2354
> > Mobile 516-967-0656
> >
> > ____________________________________________________________
> >
> > On 8/6/21, 7:41 PM, "Antibody Discussion Forum on behalf of
David
> Blum" <
> > <email obscured> on behalf of <email obscured>> wrote:
> >
> > Have you tried mag beads? We use them for histagged
proteins at
> small
> > scale but there are protein coupled beads too.
> >
> >
https://urldefense.com/v3/__https://www.thermofisher.com/order/catalog/product/88848__;!!PfbeBCCAmug!3WJ0-E0eUzj5cM-MvKLQ5OyZru8t7KhqsTPZP-Gh4ptWUlPQugH9vEtHLaTYxzoe$
> >
> >
> >
> > On Fri, Aug 6, 2021 at 6:47 PM Weis-Garcia, Frances/Sloan
> Kettering
> > Institute <email obscured>> wrote:
> >
> > > Happy Friday.
> > >
> > > Do any of you produce and purify small amounts of the new
MAbs
> you
> > make
> > > for further characterization prior to committing to a few
or
> one
> > winner?
> > > If so, how do you do this. Currently we make 350 ml
exhausted
> media
> > > (hybridoma adapted to 1% FBS prescreened for ultra-low
levels
> of
> > bovine IgG
> > > + 1 ng/ml IL6) and then purify it on a Prot G column
using and
> > FPLC. We
> > > get 1 β 8 mg, but I would like to scale this back and
simply
> it if
> > possible
> > > because we usually do not need that much.
> > >
> > > Thanks.
> > >
> > > Frances
> > >
> > >
> > > Frances Weis-Garcia, Ph.D.
> > >
> > > Head
> > > Bi-Institutional Antibody and Bioresource Core Facility
> > > (RRID: CR 017691)
> > >
> > > Associate Laboratory Member
> > > Immunology Program
> > >
> > > Memorial Sloan Kettering Cancer Center
> > > 1275 York Avenue, New York, NY 10065
> > > <
> >
>
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> > > T 646.888.2354 C 516.967.0656
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> > > ABCF-MSKCC website<
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> > > T 212.327.7030
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