Parallel immunizations of rabbits using the same antigen yield antibodies with similar, but not identical, epitopes.

A problem for the generation of polyclonal antibodies is the potential difficulties for obtaining a renewable resource due to batch-to-batch variations when the same antigen is immunized into several separate animals. Here, we have investigated this issue by determining the epitopes of antibodies ge...

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Autores principales:	Barbara Hjelm, Björn Forsström, John Löfblom, Johan Rockberg, Mathias Uhlén
Formato:	article
Lenguaje:	EN
Publicado:	Public Library of Science (PLoS) 2012
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/9794ae0f398d456291b5418631d3987c
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spelling	oai:doaj.org-article:9794ae0f398d456291b5418631d3987c2021-11-18T08:04:36ZParallel immunizations of rabbits using the same antigen yield antibodies with similar, but not identical, epitopes.1932-620310.1371/journal.pone.0045817https://doaj.org/article/9794ae0f398d456291b5418631d3987c2012-01-01T00:00:00Zhttps://www.ncbi.nlm.nih.gov/pmc/articles/pmid/23284606/?tool=EBIhttps://doaj.org/toc/1932-6203A problem for the generation of polyclonal antibodies is the potential difficulties for obtaining a renewable resource due to batch-to-batch variations when the same antigen is immunized into several separate animals. Here, we have investigated this issue by determining the epitopes of antibodies generated from parallel immunizations of rabbits with recombinant antigens corresponding to ten human protein targets. The epitopes were mapped by both a suspension bead array approach using overlapping synthetic 15-mer peptides and a bacterial display approach using expression of random fragments of the antigen on the surface of bacteria. Both methods determined antibody binding with the aid of fluorescent-based analysis. In addition, one polyclonal antibody was fractionated by peptide-specific affinity capture for in-depth comparison of epitopes. The results show that the same antigen immunized in several rabbits yields polyclonal antibodies with similar epitopes, but with larger differences in the relative amounts of antibodies to the different epitopes. In some cases, unique epitopes were observed for one of the immunizations. The results suggest that polyclonal antibodies generated by repeated immunizations do not display an identical epitope pattern, although many of the epitopes are similar.Barbara HjelmBjörn ForsströmJohn LöfblomJohan RockbergMathias UhlénPublic Library of Science (PLoS)articleMedicineRScienceQENPLoS ONE, Vol 7, Iss 12, p e45817 (2012)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Barbara Hjelm Björn Forsström John Löfblom Johan Rockberg Mathias Uhlén Parallel immunizations of rabbits using the same antigen yield antibodies with similar, but not identical, epitopes.
description	A problem for the generation of polyclonal antibodies is the potential difficulties for obtaining a renewable resource due to batch-to-batch variations when the same antigen is immunized into several separate animals. Here, we have investigated this issue by determining the epitopes of antibodies generated from parallel immunizations of rabbits with recombinant antigens corresponding to ten human protein targets. The epitopes were mapped by both a suspension bead array approach using overlapping synthetic 15-mer peptides and a bacterial display approach using expression of random fragments of the antigen on the surface of bacteria. Both methods determined antibody binding with the aid of fluorescent-based analysis. In addition, one polyclonal antibody was fractionated by peptide-specific affinity capture for in-depth comparison of epitopes. The results show that the same antigen immunized in several rabbits yields polyclonal antibodies with similar epitopes, but with larger differences in the relative amounts of antibodies to the different epitopes. In some cases, unique epitopes were observed for one of the immunizations. The results suggest that polyclonal antibodies generated by repeated immunizations do not display an identical epitope pattern, although many of the epitopes are similar.
format	article
author	Barbara Hjelm Björn Forsström John Löfblom Johan Rockberg Mathias Uhlén
author_facet	Barbara Hjelm Björn Forsström John Löfblom Johan Rockberg Mathias Uhlén
author_sort	Barbara Hjelm
title	Parallel immunizations of rabbits using the same antigen yield antibodies with similar, but not identical, epitopes.
title_short	Parallel immunizations of rabbits using the same antigen yield antibodies with similar, but not identical, epitopes.
title_full	Parallel immunizations of rabbits using the same antigen yield antibodies with similar, but not identical, epitopes.
title_fullStr	Parallel immunizations of rabbits using the same antigen yield antibodies with similar, but not identical, epitopes.
title_full_unstemmed	Parallel immunizations of rabbits using the same antigen yield antibodies with similar, but not identical, epitopes.
title_sort	parallel immunizations of rabbits using the same antigen yield antibodies with similar, but not identical, epitopes.
publisher	Public Library of Science (PLoS)
publishDate	2012
url	https://doaj.org/article/9794ae0f398d456291b5418631d3987c
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Parallel immunizations of rabbits using the same antigen yield antibodies with similar, but not identical, epitopes.

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