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Jens Christian Jensenius

A method for the blocking of endogenous immunoglobulin on frozen tissue sections in the screening of human hybridoma antibody in culture supernatants

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A method for the blocking of endogenous immunoglobulin on frozen tissue sections in the screening of human hybridoma antibody in culture supernatants. / Nielsen, B; Borup-Christensen, P; Erb, Karin; Jensenius, Jens Christian; Husby, Sofie.

In: Hybridoma, Vol. 6, No. 1, 1987, p. 103-9.

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@article{75b7c1d8b3ab49169559a1f5b76ec2f2,
title = "A method for the blocking of endogenous immunoglobulin on frozen tissue sections in the screening of human hybridoma antibody in culture supernatants",
abstract = "Endogenous immunoglobulin in tissue sections pose a problem in immuno-histochemical techniques employing homologous antibody as primary reagents and enzyme-labelled anti-immunoglobulin for the development. A method for the blocking of endogenous immunoglobulin in human tissue sections by incubation with monomeric pepsin fragments (Fab') of rabbit anti-human immunoglobulin before applying monoclonal antibody was evaluated for the screening of human monoclonal antibody. It was initially demonstrated that Fab' rabbit anti-human IgM and anti-IgG could block endogenous immunoglobulin in human IgM and IgG producing tumors thereby abolishing the binding of subsequently applied peroxidase-labelled anti-IgM or anti-IgG. Frozen sections of human colo-rectal adenocarcinomas show a variable background staining caused by the endogenous immunoglobulin. The background completely disappeared in the IgM system by preincubation with Fab' anti-IgM while the background was clearly reduced but not abolished in the IgG system. A human hybridoma supernatant containing IgM reactive with colo-rectal adenocarcinoma could easily be screened on frozen sections using this method. This approach should be generally useful for the screening of human antibody on human tissue sections.",
keywords = "Antibodies, Monoclonal, Colonic Neoplasms, Humans, Hybridomas, Immunoglobulin G, Immunoglobulin M, Immunoglobulins, Lymphoma, Plasmacytoma, Rectal Neoplasms",
author = "B Nielsen and P Borup-Christensen and Karin Erb and Jensenius, {Jens Christian} and Sofie Husby",
year = "1987",
language = "English",
volume = "6",
pages = "103--9",
journal = "Hybridoma",
issn = "1554-0014",
publisher = "Mary Ann/Liebert, Inc. Publishers",
number = "1",

}

RIS

TY - JOUR

T1 - A method for the blocking of endogenous immunoglobulin on frozen tissue sections in the screening of human hybridoma antibody in culture supernatants

AU - Nielsen, B

AU - Borup-Christensen, P

AU - Erb, Karin

AU - Jensenius, Jens Christian

AU - Husby, Sofie

PY - 1987

Y1 - 1987

N2 - Endogenous immunoglobulin in tissue sections pose a problem in immuno-histochemical techniques employing homologous antibody as primary reagents and enzyme-labelled anti-immunoglobulin for the development. A method for the blocking of endogenous immunoglobulin in human tissue sections by incubation with monomeric pepsin fragments (Fab') of rabbit anti-human immunoglobulin before applying monoclonal antibody was evaluated for the screening of human monoclonal antibody. It was initially demonstrated that Fab' rabbit anti-human IgM and anti-IgG could block endogenous immunoglobulin in human IgM and IgG producing tumors thereby abolishing the binding of subsequently applied peroxidase-labelled anti-IgM or anti-IgG. Frozen sections of human colo-rectal adenocarcinomas show a variable background staining caused by the endogenous immunoglobulin. The background completely disappeared in the IgM system by preincubation with Fab' anti-IgM while the background was clearly reduced but not abolished in the IgG system. A human hybridoma supernatant containing IgM reactive with colo-rectal adenocarcinoma could easily be screened on frozen sections using this method. This approach should be generally useful for the screening of human antibody on human tissue sections.

AB - Endogenous immunoglobulin in tissue sections pose a problem in immuno-histochemical techniques employing homologous antibody as primary reagents and enzyme-labelled anti-immunoglobulin for the development. A method for the blocking of endogenous immunoglobulin in human tissue sections by incubation with monomeric pepsin fragments (Fab') of rabbit anti-human immunoglobulin before applying monoclonal antibody was evaluated for the screening of human monoclonal antibody. It was initially demonstrated that Fab' rabbit anti-human IgM and anti-IgG could block endogenous immunoglobulin in human IgM and IgG producing tumors thereby abolishing the binding of subsequently applied peroxidase-labelled anti-IgM or anti-IgG. Frozen sections of human colo-rectal adenocarcinomas show a variable background staining caused by the endogenous immunoglobulin. The background completely disappeared in the IgM system by preincubation with Fab' anti-IgM while the background was clearly reduced but not abolished in the IgG system. A human hybridoma supernatant containing IgM reactive with colo-rectal adenocarcinoma could easily be screened on frozen sections using this method. This approach should be generally useful for the screening of human antibody on human tissue sections.

KW - Antibodies, Monoclonal

KW - Colonic Neoplasms

KW - Humans

KW - Hybridomas

KW - Immunoglobulin G

KW - Immunoglobulin M

KW - Immunoglobulins

KW - Lymphoma

KW - Plasmacytoma

KW - Rectal Neoplasms

M3 - Journal article

C2 - 3679252

VL - 6

SP - 103

EP - 109

JO - Hybridoma

JF - Hybridoma

SN - 1554-0014

IS - 1

ER -