A novel affinity purification method to isolate peptide specific antibodies
Karlsen, Allan ; Lernmark, Åke LU
; Kofod, Hans
and Dyrberg, Thomas
(1990)
In Journal of Immunological Methods
128(2).
p.151-157
- Abstract
Site-specific, high affinity polyclonal antisera are effectively and successfully produced by immunizing rabbits with synthetic peptides. The use of these antisera in subsequent immune analysis is often limited because of non-specific binding. We describe a new and simple method to effectively affinity-purify anti-peptide antibodies. To test our system, rabbits were immunized with model peptides representing sequences of the putative rabbit growth hormone receptor and several HLA-DQ β-chain molecules. Polystyrene plastic beads were coated with peptides. Immune serum was incubated with the beads and after a wash step the bound antibodies were eluted in 1 M acetic acid. The eluted material was composed predominantly of intact... (More)
Site-specific, high affinity polyclonal antisera are effectively and successfully produced by immunizing rabbits with synthetic peptides. The use of these antisera in subsequent immune analysis is often limited because of non-specific binding. We describe a new and simple method to effectively affinity-purify anti-peptide antibodies. To test our system, rabbits were immunized with model peptides representing sequences of the putative rabbit growth hormone receptor and several HLA-DQ β-chain molecules. Polystyrene plastic beads were coated with peptides. Immune serum was incubated with the beads and after a wash step the bound antibodies were eluted in 1 M acetic acid. The eluted material was composed predominantly of intact immunoglobulin as evidenced by the presence of heavy and light chain bands in SDS-PAGE. The eluted antibodies were peptide specific in ELISA and bound only to intact, antigenic protein in immunoblot analyses. The sequence-specific nature of the eluted antibodies was confirmed since binding to the antigenic proteins could be displayed by the immunizing but not by unrelated peptides.
(Less)
- author
- Karlsen, Allan
; Lernmark, Åke
LU
; Kofod, Hans
and Dyrberg, Thomas
- publishing date
- 1990-04-17
- type
- Contribution to journal
- publication status
- published
- keywords
- Affinity purification, Anti-peptide antibody, Growth hormone receptor, HLA class II β chain molecule
- in
- Journal of Immunological Methods
- volume
- 128
- issue
- 2
- pages
- 151 - 157
- publisher
- Elsevier
- external identifiers
-
- scopus:0025373281
- pmid:2324508
- ISSN
- 0022-1759
- DOI
- 10.1016/0022-1759(90)90205-A
- language
- English
- LU publication?
- no
- id
- b3b420f8-2a58-4795-82c3-86e03a25d55b
- date added to LUP
- 2019-09-11 09:42:28
- date last changed
- 2024-03-13 08:10:10
@article{b3b420f8-2a58-4795-82c3-86e03a25d55b,
abstract = {{<p>Site-specific, high affinity polyclonal antisera are effectively and successfully produced by immunizing rabbits with synthetic peptides. The use of these antisera in subsequent immune analysis is often limited because of non-specific binding. We describe a new and simple method to effectively affinity-purify anti-peptide antibodies. To test our system, rabbits were immunized with model peptides representing sequences of the putative rabbit growth hormone receptor and several HLA-DQ β-chain molecules. Polystyrene plastic beads were coated with peptides. Immune serum was incubated with the beads and after a wash step the bound antibodies were eluted in 1 M acetic acid. The eluted material was composed predominantly of intact immunoglobulin as evidenced by the presence of heavy and light chain bands in SDS-PAGE. The eluted antibodies were peptide specific in ELISA and bound only to intact, antigenic protein in immunoblot analyses. The sequence-specific nature of the eluted antibodies was confirmed since binding to the antigenic proteins could be displayed by the immunizing but not by unrelated peptides.</p>}},
author = {{Karlsen, Allan and Lernmark, Åke and Kofod, Hans and Dyrberg, Thomas}},
issn = {{0022-1759}},
keywords = {{Affinity purification; Anti-peptide antibody; Growth hormone receptor; HLA class II β chain molecule}},
language = {{eng}},
month = {{04}},
number = {{2}},
pages = {{151--157}},
publisher = {{Elsevier}},
series = {{Journal of Immunological Methods}},
title = {{A novel affinity purification method to isolate peptide specific antibodies}},
url = {{http://dx.doi.org/10.1016/0022-1759(90)90205-A}},
doi = {{10.1016/0022-1759(90)90205-A}},
volume = {{128}},
year = {{1990}},
}