|
rdf:type |
|
|
lifeskim:mentions |
|
|
pubmed:issue |
1
|
|
pubmed:dateCreated |
2000-2-29
|
|
pubmed:abstractText |
The creation of large phage antibody libraries has become an important goal in selecting antibodies against any antigen. Here we describe a method for making libraries so large that the complete diversity cannot be accessed using traditional phage technology. This involves the creation of a primary phage scFv library in a phagemid vector containing two nonhomologous lox sites. Contrary to the current dogma, we found that infecting Cre recombinase-expressing bacteria by such a primary library at a high multiplicity of infection results in the entry of many different phagemid into the cell. Exchange of Vh and Vl genes between such phagemids creates many new V h/Vl combinations, all of which are functional. On the basis of the observed recombination, the library is calculated to have a diversity of 3x1011. A library created using this method was validated by the selection of high affinity antibodies against a large number of different protein antigens.
|
|
pubmed:language |
eng
|
|
pubmed:journal |
|
|
pubmed:citationSubset |
IM
|
|
pubmed:chemical |
|
|
pubmed:status |
MEDLINE
|
|
pubmed:month |
Jan
|
|
pubmed:issn |
1087-0156
|
|
pubmed:author |
|
|
pubmed:issnType |
Print
|
|
pubmed:volume |
18
|
|
pubmed:owner |
NLM
|
|
pubmed:authorsComplete |
Y
|
|
pubmed:pagination |
75-80
|
|
pubmed:dateRevised |
2011-11-17
|
|
pubmed:meshHeading |
pubmed-meshheading:10625396-Antibodies,
pubmed-meshheading:10625396-Antibody Diversity,
pubmed-meshheading:10625396-Antibody Specificity,
pubmed-meshheading:10625396-Antigens,
pubmed-meshheading:10625396-Attachment Sites, Microbiological,
pubmed-meshheading:10625396-Bacteriophages,
pubmed-meshheading:10625396-Binding Sites, Antibody,
pubmed-meshheading:10625396-Cloning, Molecular,
pubmed-meshheading:10625396-Escherichia coli,
pubmed-meshheading:10625396-Gene Rearrangement,
pubmed-meshheading:10625396-Genetic Vectors,
pubmed-meshheading:10625396-Humans,
pubmed-meshheading:10625396-Immunoglobulin Fragments,
pubmed-meshheading:10625396-Integrases,
pubmed-meshheading:10625396-Peptide Library,
pubmed-meshheading:10625396-Recombination, Genetic,
pubmed-meshheading:10625396-Reproducibility of Results,
pubmed-meshheading:10625396-Sequence Analysis, DNA,
pubmed-meshheading:10625396-Viral Proteins
|
|
pubmed:year |
2000
|
|
pubmed:articleTitle |
Exploiting recombination in single bacteria to make large phage antibody libraries.
|
|
pubmed:affiliation |
International School for Advanced Studies (SISSA), Biophysics Sector, Via Beirut 2-4, Trieste, 34014, Italy.
|
|
pubmed:publicationType |
Journal Article,
Research Support, Non-U.S. Gov't
|
