A novel affinity purification method to isolate peptide specific antibodies

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A novel affinity purification method to isolate peptide specific antibodies. / Karlsen, Alan E; Lernmark, A; Kofod, Hans; Dyrberg, T.

In: Journal of Immunological Methods, Vol. 128, No. 2, 17.04.1990, p. 151-7.

Research output: Contribution to journal › Journal article › Research › peer-review

Harvard

Karlsen, AE, Lernmark, A, Kofod, H & Dyrberg, T 1990, 'A novel affinity purification method to isolate peptide specific antibodies', Journal of Immunological Methods, vol. 128, no. 2, pp. 151-7.

APA

Karlsen, A. E., Lernmark, A., Kofod, H., & Dyrberg, T. (1990). A novel affinity purification method to isolate peptide specific antibodies. Journal of Immunological Methods, 128(2), 151-7.

Vancouver

Karlsen AE, Lernmark A, Kofod H, Dyrberg T. A novel affinity purification method to isolate peptide specific antibodies. Journal of Immunological Methods. 1990 Apr 17;128(2):151-7.

Author

Karlsen, Alan E ; Lernmark, A ; Kofod, Hans ; Dyrberg, T. / A novel affinity purification method to isolate peptide specific antibodies. In: Journal of Immunological Methods. 1990 ; Vol. 128, No. 2. pp. 151-7.

Bibtex

@article{4ed2bb90bd444c978c6bcc13893f91e9,
title = "A novel affinity purification method to isolate peptide specific antibodies",
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 beta-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 displaced by the immunizing but not by unrelated peptides.",
keywords = "Animals, Antibodies, Antibody Specificity, Chromatography, Affinity, Electrophoresis, Polyacrylamide Gel, Enzyme-Linked Immunosorbent Assay, HLA-DQ Antigens, Immunoblotting, Peptides, Rabbits, Rats, Receptors, Somatotropin",
author = "Karlsen, {Alan E} and A Lernmark and Hans Kofod and T Dyrberg",
year = "1990",
month = apr,
day = "17",
language = "English",
volume = "128",
pages = "151--7",
journal = "Journal of Immunological Methods",
issn = "0022-1759",
publisher = "Elsevier",
number = "2",

}

RIS

TY - JOUR

T1 - A novel affinity purification method to isolate peptide specific antibodies

AU - Karlsen, Alan E

AU - Lernmark, A

AU - Kofod, Hans

AU - Dyrberg, T

PY - 1990/4/17

Y1 - 1990/4/17

N2 - 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 beta-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 displaced by the immunizing but not by unrelated peptides.

AB - 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 beta-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 displaced by the immunizing but not by unrelated peptides.

KW - Animals

KW - Antibodies

KW - Antibody Specificity

KW - Chromatography, Affinity

KW - Electrophoresis, Polyacrylamide Gel

KW - Enzyme-Linked Immunosorbent Assay

KW - HLA-DQ Antigens

KW - Immunoblotting

KW - Peptides

KW - Rabbits

KW - Rats

KW - Receptors, Somatotropin

M3 - Journal article

C2 - 2324508

VL - 128

SP - 151

EP - 157

JO - Journal of Immunological Methods

JF - Journal of Immunological Methods

SN - 0022-1759

IS - 2

ER -

ID: 45574812