Selection of phage displayed antibodies based on kinetic constants
Dueñas, Marta ; Malmborg, Ann-Christin LU ; Casalvilla, Racmar ; Ohlin, Mats LU
and Borrebaeck, C. A K
LU
(1996)
In Molecular Immunology
33(3).
p.279-285
- Abstract
The display of antibody fragments on the surface of filamentous bacteriophages and the selection of binders from antibody libraries have provided powerful tools to generate human antibodies. We reported recently a new concept (SAP system) for the selection of specific phages by linking antigenic recognition and phage replication, using a soluble fusion protein containing the antigen and a fragment of the M13 coat protein 3. In this investigation, a model library has been composed using six different antibody fragments which were characterized individually regarding their k(ass), k(diss) and K(a). All Feb fragments were specific for a 15 amino acid region of the V3 loop of gp120 (HIV-1). We demonstrated that the SAP system could... (More)
The display of antibody fragments on the surface of filamentous bacteriophages and the selection of binders from antibody libraries have provided powerful tools to generate human antibodies. We reported recently a new concept (SAP system) for the selection of specific phages by linking antigenic recognition and phage replication, using a soluble fusion protein containing the antigen and a fragment of the M13 coat protein 3. In this investigation, a model library has been composed using six different antibody fragments which were characterized individually regarding their k(ass), k(diss) and K(a). All Feb fragments were specific for a 15 amino acid region of the V3 loop of gp120 (HIV-1). We demonstrated that the SAP system could discriminate between the kinetic parameters of each clone, using different selection strategies. Phages expressing high affinity clones were selected preferentially using low doses of antigen but clones of lower affinity also could be selected by increasing the antigen concentration or using a preselection procedure. Phages expressing antibody fragment with high association or low dissociation rate constants were retrieved by utilizing short contact times between antigen and antibody or antigen-chase conditions.
(Less)
- author
- Dueñas, Marta
; Malmborg, Ann-Christin
LU
; Casalvilla, Racmar
; Ohlin, Mats
LU
and Borrebaeck, C. A K
LU
- organization
- publishing date
- 1996-02
- type
- Contribution to journal
- publication status
- published
- keywords
- Antibody, Antigen-specific selection, Association, Dissociation, Phage display, Rate constant, SAP
- in
- Molecular Immunology
- volume
- 33
- issue
- 3
- pages
- 7 pages
- publisher
- Pergamon Press Ltd.
- external identifiers
-
- pmid:8649449
- scopus:0029901468
- ISSN
- 0161-5890
- DOI
- 10.1016/0161-5890(95)00137-9
- language
- English
- LU publication?
- yes
- id
- 69a48a39-61ee-4f0f-ae17-1383d37f49b2
- date added to LUP
- 2016-04-19 14:10:54
- date last changed
- 2024-01-04 02:22:41
@article{69a48a39-61ee-4f0f-ae17-1383d37f49b2,
abstract = {{<p>The display of antibody fragments on the surface of filamentous bacteriophages and the selection of binders from antibody libraries have provided powerful tools to generate human antibodies. We reported recently a new concept (SAP system) for the selection of specific phages by linking antigenic recognition and phage replication, using a soluble fusion protein containing the antigen and a fragment of the M13 coat protein 3. In this investigation, a model library has been composed using six different antibody fragments which were characterized individually regarding their k(ass), k(diss) and K(a). All Feb fragments were specific for a 15 amino acid region of the V3 loop of gp120 (HIV-1). We demonstrated that the SAP system could discriminate between the kinetic parameters of each clone, using different selection strategies. Phages expressing high affinity clones were selected preferentially using low doses of antigen but clones of lower affinity also could be selected by increasing the antigen concentration or using a preselection procedure. Phages expressing antibody fragment with high association or low dissociation rate constants were retrieved by utilizing short contact times between antigen and antibody or antigen-chase conditions.</p>}},
author = {{Dueñas, Marta and Malmborg, Ann-Christin and Casalvilla, Racmar and Ohlin, Mats and Borrebaeck, C. A K}},
issn = {{0161-5890}},
keywords = {{Antibody; Antigen-specific selection; Association; Dissociation; Phage display; Rate constant; SAP}},
language = {{eng}},
number = {{3}},
pages = {{279--285}},
publisher = {{Pergamon Press Ltd.}},
series = {{Molecular Immunology}},
title = {{Selection of phage displayed antibodies based on kinetic constants}},
url = {{http://dx.doi.org/10.1016/0161-5890(95)00137-9}},
doi = {{10.1016/0161-5890(95)00137-9}},
volume = {{33}},
year = {{1996}},
}