🤔.
Dream a little dream on this.
Laura Bover, PhD
(working from home during COVID-19 Cell 713-703-1951<tel:713-703-1951>)
Professor
Director Monoclonal Antibody Core Facility
University of Texas M.D.Anderson Cancer Center
Associate member of The University of Texas Graduate School of Biomedical
Sciences at Houston
Member of the Scientific Review Committee 2 (former Clinical Research Committee
2)
Member of the Steering Committee of the Immunology Program GSBS
Immunology Department/ Genomics Medicine Department
7455 Fannin St<x-apple-data-detectors://2/1> Room 1SCR4.2021
Houston- TX 77054<x-apple-data-detectors://3/0>
T 713-563-3301<tel:713-563-3301>
F 713-563-3276<tel:713-563-3276>
<email obscured><email obscured>>
In order to continue obtaining Core support in a challenging environment, it is
crucial that our users acknowledge the following grants in all publications
(see following text). Thank you!
“We acknowledge the UT-MDACC Monoclonal Antibodies Core facility, supported in
part by CCSG grant number P30 CA016672.”
Sent from my iPhone
On Jul 27, 2021, at 10:32 PM, Weis-Garcia, Frances/Sloan Kettering Institute
<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.
Happy Wednesday everyone!
We have an electrofusion where half the antigen specific hybridomas we have
secrete both IgG and IgG base on 2 sandwich ELISAs:
* IgG ELISA
* Capture goat anti-mouse Fc gamma specific
* Detect goat anti-mouse F(Ab’)2
* IgM ELISA
* Capture goat anti-mouse IgM, mu specific
* Detect goat anti-mouse F(Ab’)2
The other half only secrete IgG.
The only thing I can think of the hybridomas have 2 B cells – one IgG and 1
IgM. We have not subcloned yet … but these cultures did come from a ClonePix
run.
Thoughts ???
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
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!2RS2q-1wTYHGfxNvvpvsV2BCD9o8krF3Ar0LVMpHhkHUfkv_MUbCxbUnaO_qUuai$
>
Adjunct Faculty
Center for Clinical and Translational Science
The Rockefeller University
1230 York Avenue, New York, NY 10065
T 212.327.7030
ABCF-RU
website<https://urldefense.com/v3/__https://www.rockefeller.edu/monoclonal/__;!!PfbeBCCAmug!2RS2q-1wTYHGfxNvvpvsV2BCD9o8krF3Ar0LVMpHhkHUfkv_MUbCxbUnaFYFUkAw$
>
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Rest of post
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Thanks for ponding this with me.
I think we are going to IP the media with Protein L beads and see what to get
to confirm the ELISA results. Next set would be to subclone a few to make sure
they antibodies are coming from one cell and they are not mixed cultures.
Frances
ZRC 1563
Office 646-888-2354
Mobile 516-967-0656
On 7/28/21, 12:25 AM, "Antibody Discussion Forum on behalf of Bover,Laura"
<email obscured> on behalf of <email obscured>> wrote:
🤔.
Dream a little dream on this.
Laura Bover, PhD
(working from home during COVID-19 Cell 713-703-1951<tel:713-703-1951>)
Professor
Director Monoclonal Antibody Core Facility
University of Texas M.D.Anderson Cancer Center
Associate member of The University of Texas Graduate School of Biomedical
Sciences at Houston
Member of the Scientific Review Committee 2 (former Clinical Research
Committee 2)
Member of the Steering Committee of the Immunology Program GSBS
Immunology Department/ Genomics Medicine Department
7455 Fannin St<x-apple-data-detectors://2/1> Room 1SCR4.2021
Houston- TX 77054<x-apple-data-detectors://3/0>
T 713-563-3301<tel:713-563-3301>
F 713-563-3276<tel:713-563-3276>
<email obscured><email obscured>>
In order to continue obtaining Core support in a challenging environment,
it is crucial that our users acknowledge the following grants in all
publications (see following text). Thank you!
“We acknowledge the UT-MDACC Monoclonal Antibodies Core facility,
supported in part by CCSG grant number P30 CA016672.”
Sent from my iPhone
On Jul 27, 2021, at 10:32 PM, Weis-Garcia, Frances/Sloan Kettering
Institute <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.
Happy Wednesday everyone!
We have an electrofusion where half the antigen specific hybridomas we have
secrete both IgG and IgG base on 2 sandwich ELISAs:
* IgG ELISA
* Capture goat anti-mouse Fc gamma specific
* Detect goat anti-mouse F(Ab’)2
* IgM ELISA
* Capture goat anti-mouse IgM, mu specific
* Detect goat anti-mouse F(Ab’)2
The other half only secrete IgG.
The only thing I can think of the hybridomas have 2 B cells – one IgG and 1
IgM. We have not subcloned yet … but these cultures did come from a ClonePix
run.
Thoughts ???
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
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!2RS2q-1wTYHGfxNvvpvsV2BCD9o8krF3Ar0LVMpHhkHUfkv_MUbCxbUnaO_qUuai$
>
Adjunct Faculty
Center for Clinical and Translational Science
The Rockefeller University
1230 York Avenue, New York, NY 10065
T 212.327.7030
ABCF-RU
website<https://urldefense.com/v3/__https://www.rockefeller.edu/monoclonal/__;!!PfbeBCCAmug!2RS2q-1wTYHGfxNvvpvsV2BCD9o8krF3Ar0LVMpHhkHUfkv_MUbCxbUnaFYFUkAw$
>
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I think this is the way to go. Please tell us the end of the saga.
Laura Bover
Sent from my iPhone
On Jul 28, 2021, at 10:57 AM, Weis-Garcia, Frances/Sloan Kettering Institute
<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.
Thanks for ponding this with me.
I think we are going to IP the media with Protein L beads and see what to get
to confirm the ELISA results. Next set would be to subclone a few to make sure
they antibodies are coming from one cell and they are not mixed cultures.
Frances
ZRC 1563
Office 646-888-2354
Mobile 516-967-0656
On 7/28/21, 12:25 AM, "Antibody Discussion Forum on behalf of Bover,Laura"
<email obscured> on behalf of <email obscured>> wrote:
🤔.
Dream a little dream on this.
Laura Bover, PhD
(working from home during COVID-19 Cell 713-703-1951<tel:713-703-1951>)
Professor
Director Monoclonal Antibody Core Facility
University of Texas M.D.Anderson Cancer Center
Associate member of The University of Texas Graduate School of Biomedical
Sciences at Houston
Member of the Scientific Review Committee 2 (former Clinical Research
Committee 2)
Member of the Steering Committee of the Immunology Program GSBS
Immunology Department/ Genomics Medicine Department
7455 Fannin St<x-apple-data-detectors://2/1> Room 1SCR4.2021
Houston- TX 77054<x-apple-data-detectors://3/0>
T 713-563-3301<tel:713-563-3301>
F 713-563-3276<tel:713-563-3276>
<email obscured><email obscured>>
In order to continue obtaining Core support in a challenging environment, it
is crucial that our users acknowledge the following grants in all publications
(see following text). Thank you!
“We acknowledge the UT-MDACC Monoclonal Antibodies Core facility, supported
in part by CCSG grant number P30 CA016672.”
Sent from my iPhone
On Jul 27, 2021, at 10:32 PM, Weis-Garcia, Frances/Sloan Kettering Institute
<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.
Happy Wednesday everyone!
We have an electrofusion where half the antigen specific hybridomas we have
secrete both IgG and IgG base on 2 sandwich ELISAs:
* IgG ELISA
* Capture goat anti-mouse Fc gamma specific
* Detect goat anti-mouse F(Ab’)2
* IgM ELISA
* Capture goat anti-mouse IgM, mu specific
* Detect goat anti-mouse F(Ab’)2
The other half only secrete IgG.
The only thing I can think of the hybridomas have 2 B cells – one IgG and 1
IgM. We have not subcloned yet … but these cultures did come from a ClonePix
run.
Thoughts ???
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
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!2RS2q-1wTYHGfxNvvpvsV2BCD9o8krF3Ar0LVMpHhkHUfkv_MUbCxbUnaO_qUuai$
>
Adjunct Faculty
Center for Clinical and Translational Science
The Rockefeller University
1230 York Avenue, New York, NY 10065
T 212.327.7030
ABCF-RU
website<https://urldefense.com/v3/__https://www.rockefeller.edu/monoclonal/__;!!PfbeBCCAmug!2RS2q-1wTYHGfxNvvpvsV2BCD9o8krF3Ar0LVMpHhkHUfkv_MUbCxbUnaFYFUkAw$
>
Please note that this e-mail and any files transmitted with it may be
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I guess my only two cents is we had way better luck with capture select resins
than Protein L. Can get kappa or lambda specific resins.
Good luck.
Rob
From: Antibody Discussion Forum <email obscured>> on behalf of
Bover,Laura <email obscured>>
Date: Thursday, July 29, 2021 at 8:43 AM
To: <email obscured> <email obscured>>
Subject: Re: [ABRF Ab Forum] Re: [EXT] Hybridoma sups scoring positive for IgG
and IgM
I think this is the way to go. Please tell us the end of the saga.
Laura Bover
Sent from my iPhone
On Jul 28, 2021, at 10:57 AM, Weis-Garcia, Frances/Sloan Kettering Institute
<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.
Thanks for ponding this with me.
I think we are going to IP the media with Protein L beads and see what to get
to confirm the ELISA results. Next set would be to subclone a few to make sure
they antibodies are coming from one cell and they are not mixed cultures.
Frances
ZRC 1563
Office 646-888-2354
Mobile 516-967-0656
On 7/28/21, 12:25 AM, "Antibody Discussion Forum on behalf of Bover,Laura"
<email obscured> on behalf of <email obscured>> wrote:
🤔.
Dream a little dream on this.
Laura Bover, PhD
(working from home during COVID-19 Cell 713-703-1951<tel:713-703-1951>)
Professor
Director Monoclonal Antibody Core Facility
University of Texas M.D.Anderson Cancer Center
Associate member of The University of Texas Graduate School of Biomedical
Sciences at Houston
Member of the Scientific Review Committee 2 (former Clinical Research
Committee 2)
Member of the Steering Committee of the Immunology Program GSBS
Immunology Department/ Genomics Medicine Department
7455 Fannin St<x-apple-data-detectors://2/1> Room 1SCR4.2021
Houston- TX 77054<x-apple-data-detectors://3/0>
T 713-563-3301<tel:713-563-3301>
F 713-563-3276<tel:713-563-3276>
<email obscured><email obscured>>
In order to continue obtaining Core support in a challenging environment, it
is crucial that our users acknowledge the following grants in all publications
(see following text). Thank you!
“We acknowledge the UT-MDACC Monoclonal Antibodies Core facility, supported
in part by CCSG grant number P30 CA016672.”
Sent from my iPhone
On Jul 27, 2021, at 10:32 PM, Weis-Garcia, Frances/Sloan Kettering Institute
<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.
Happy Wednesday everyone!
We have an electrofusion where half the antigen specific hybridomas we have
secrete both IgG and IgG base on 2 sandwich ELISAs:
* IgG ELISA
* Capture goat anti-mouse Fc gamma specific
* Detect goat anti-mouse F(Ab’)2
* IgM ELISA
* Capture goat anti-mouse IgM, mu specific
* Detect goat anti-mouse F(Ab’)2
The other half only secrete IgG.
The only thing I can think of the hybridomas have 2 B cells – one IgG and 1
IgM. We have not subcloned yet … but these cultures did come from a ClonePix
run.
Thoughts ???
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
T 646.888.2354 C 516.967.0656
ABCF-MSKCC
website<https://nam04.safelinks.protection.outlook.com/?url=https%3A%2F%2Furldefense.com%2Fv3%2F__https%3A%2F%2Fwww.mskcc.org%2Fresearch-advantage%2Fcore-facilities%2Fmonoclonal-antibody-core-facility__%3B!!PfbeBCCAmug!2RS2q-1wTYHGfxNvvpvsV2BCD9o8krF3Ar0LVMpHhkHUfkv_MUbCxbUnaO_qUuai%24&data=04%7C01%7Crobert.carnahan%40vanderbilt.edu%7Cf0b6fb49e42b4e92aaf708d95296e5ff%7Cba5a7f39e3be4ab3b45067fa80faecad%7C0%7C1%7C637631630326968143%7CUnknown%7CTWFpbGZsb3d8eyJWIjoiMC4wLjAwMDAiLCJQIjoiV2luMzIiLCJBTiI6Ik1haWwiLCJXVCI6Mn0%3D%7C1000&sdata=BxaTRTc2GKSCdyN%2Be4%2FLdzNhZY6Bv3iYM2YHDkTA6ho%3D&reserved=0
>
Adjunct Faculty
Center for Clinical and Translational Science
The Rockefeller University
1230 York Avenue, New York, NY 10065
T 212.327.7030
ABCF-RU
website<https://nam04.safelinks.protection.outlook.com/?url=https%3A%2F%2Furldefense.com%2Fv3%2F__https%3A%2F%2Fwww.rockefeller.edu%2Fmonoclonal%2F__%3B!!PfbeBCCAmug!2RS2q-1wTYHGfxNvvpvsV2BCD9o8krF3Ar0LVMpHhkHUfkv_MUbCxbUnaFYFUkAw%24&data=04%7C01%7Crobert.carnahan%40vanderbilt.edu%7Cf0b6fb49e42b4e92aaf708d95296e5ff%7Cba5a7f39e3be4ab3b45067fa80faecad%7C0%7C1%7C637631630326968143%7CUnknown%7CTWFpbGZsb3d8eyJWIjoiMC4wLjAwMDAiLCJQIjoiV2luMzIiLCJBTiI6Ik1haWwiLCJXVCI6Mn0%3D%7C1000&sdata=CFmrwdN3PYMmZvtVNnor43T%2Bcwu8n%2BW4b0wLSGZutkM%3D&reserved=0
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I agree, if we were purifying.
All we want to see is if they are both there with a different technique ... and
I want to SEE the HC band ... not just color in a well __
Frances
ZRC 1563
Office 646-888-2354
Mobile 516-967-0656
On 7/29/21, 9:51 AM, "Antibody Discussion Forum on behalf of Carnahan, Robert"
<email obscured> on behalf of <email obscured>> wrote:
I guess my only two cents is we had way better luck with capture select
resins than Protein L. Can get kappa or lambda specific resins.
Good luck.
Rob
From: Antibody Discussion Forum <email obscured>> on behalf of
Bover,Laura <email obscured>>
Date: Thursday, July 29, 2021 at 8:43 AM
To: <email obscured> <email obscured>>
Subject: Re: [ABRF Ab Forum] Re: [EXT] Hybridoma sups scoring positive for
IgG and IgM
I think this is the way to go. Please tell us the end of the saga.
Laura Bover
Sent from my iPhone
On Jul 28, 2021, at 10:57 AM, Weis-Garcia, Frances/Sloan Kettering
Institute <email obscured>> wrote:
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Thanks for ponding this with me.
I think we are going to IP the media with Protein L beads and see what to
get to confirm the ELISA results. Next set would be to subclone a few to make
sure they antibodies are coming from one cell and they are not mixed cultures.
Frances
ZRC 1563
Office 646-888-2354
Mobile 516-967-0656
On 7/28/21, 12:25 AM, "Antibody Discussion Forum on behalf of Bover,Laura"
<email obscured> on behalf of <email obscured>> wrote:
🤔.
Dream a little dream on this.
Laura Bover, PhD
(working from home during COVID-19 Cell 713-703-1951<tel:713-703-1951>)
Professor
Director Monoclonal Antibody Core Facility
University of Texas M.D.Anderson Cancer Center
Associate member of The University of Texas Graduate School of
Biomedical Sciences at Houston
Member of the Scientific Review Committee 2 (former Clinical Research
Committee 2)
Member of the Steering Committee of the Immunology Program GSBS
Immunology Department/ Genomics Medicine Department
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Houston- TX 77054<x-apple-data-detectors://3/0>
T 713-563-3301<tel:713-563-3301>
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<email obscured><email obscured>>
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Sent from my iPhone
On Jul 27, 2021, at 10:32 PM, Weis-Garcia, Frances/Sloan Kettering
Institute <email obscured>> wrote:
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Happy Wednesday everyone!
We have an electrofusion where half the antigen specific hybridomas we
have secrete both IgG and IgG base on 2 sandwich ELISAs:
* IgG ELISA
* Capture goat anti-mouse Fc gamma specific
* Detect goat anti-mouse F(Ab’)2
* IgM ELISA
* Capture goat anti-mouse IgM, mu specific
* Detect goat anti-mouse F(Ab’)2
The other half only secrete IgG.
The only thing I can think of the hybridomas have 2 B cells – one IgG
and 1 IgM. We have not subcloned yet … but these cultures did come from a
ClonePix run.
Thoughts ???
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
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