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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.

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author
organization
publishing date
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
external identifiers
  • 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
2016-04-22 13:22:04
@misc{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},
  keyword      = {Antibody,Antigen-specific selection,Association,Dissociation,Phage display,Rate constant,SAP},
  language     = {eng},
  number       = {3},
  pages        = {279--285},
  publisher    = {ARRAY(0x90d6da0)},
  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},
  volume       = {33},
  year         = {1996},
}