Linked Life Data

12210838

Source:http://linkedlifedata.com/resource/pubmed/id/12210838

Statements in which the resource exists as a subject.
PredicateObject
rdf:type
lifeskim:mentions
pubmed:issue
4
pubmed:dateCreated
2002-9-4
pubmed:abstractText
The neural cell adhesion molecule CHL1 (close homolog of L1) plays important roles in neurite outgrowth and neuronal survival in vitro. Reproducible and functionally active CHL1 antibodies are critical for a better understanding of the functional properties of CHL1 in vitro and in vivo. We have isolated human single-chain variable fragment (scFv) antibodies against mouse CHL1 from a human synthetic phage display library. To improve the binding activity of such antibodies, a clone (C12) was selected for affinity maturation by combined random mutagenesis of the V(H) gene and site-directed cassette mutagenesis to introduce random mutations in the complementarity determining region 3 (CDR3) of the V(L) gene. From the mutant phage display library, we selected a clone (6C2) that gave the strongest signal as determined by ELISA. The dissociation constant of 6C2 (Kd 2.28 x 10(-8) M) was increased approximately 85-fold compared with the wild-type clone C12 (Kd 1.93 x 10(-6) M). 6C2 detected CHL1 by Western blot analysis in mouse brain homogenates and detected CHL1 in CHL1-transfected cells by immunofluorescence. Furthermore, the wild-type and affinity-matured antibodies promoted neurite outgrowth of hippocampal and cerebellar neurons in vitro. Our results suggest that the affinity-matured CHL1 scFv antibody will serve a range of applications in vitro and in vivo.
pubmed:language
eng
pubmed:journal
pubmed:citationSubset
IM
pubmed:chemical
pubmed:status
MEDLINE
pubmed:month
Aug
pubmed:issn
0360-4012
pubmed:author
pubmed:copyrightInfo
Copyright 2002 Wiley-Liss, Inc.
pubmed:issnType
Print
pubmed:day
15
pubmed:volume
69
pubmed:owner
NLM
pubmed:authorsComplete
Y
pubmed:pagination
437-47
pubmed:dateRevised
2006-11-15
pubmed:meshHeading
pubmed-meshheading:12210838-Amino Acid Sequence, pubmed-meshheading:12210838-Animals, pubmed-meshheading:12210838-Antibodies, pubmed-meshheading:12210838-Antibody Specificity, pubmed-meshheading:12210838-Base Sequence, pubmed-meshheading:12210838-Cell Adhesion Molecules, pubmed-meshheading:12210838-Cell Differentiation, pubmed-meshheading:12210838-Cells, Cultured, pubmed-meshheading:12210838-Central Nervous System, pubmed-meshheading:12210838-Cerebellar Cortex, pubmed-meshheading:12210838-DNA, Complementary, pubmed-meshheading:12210838-Fluorescent Antibody Technique, pubmed-meshheading:12210838-Hippocampus, pubmed-meshheading:12210838-Membrane Proteins, pubmed-meshheading:12210838-Mice, pubmed-meshheading:12210838-Mice, Transgenic, pubmed-meshheading:12210838-Molecular Sequence Data, pubmed-meshheading:12210838-Mutagenesis, Site-Directed, pubmed-meshheading:12210838-Mutation, pubmed-meshheading:12210838-Neural Cell Adhesion Molecule L1, pubmed-meshheading:12210838-Neural Cell Adhesion Molecules, pubmed-meshheading:12210838-Neurites, pubmed-meshheading:12210838-Peptide Fragments, pubmed-meshheading:12210838-Peptide Library, pubmed-meshheading:12210838-Protein Structure, Tertiary, pubmed-meshheading:12210838-Proteins
pubmed:year
2002
pubmed:articleTitle
Single-chain variable fragment antibodies against the neural adhesion molecule CHL1 (close homolog of L1) enhance neurite outgrowth.
pubmed:affiliation
Zentrum für Molekulare Neurobiologie, Universität Hamburg, Germany.
pubmed:publicationType
Journal Article, Research Support, Non-U.S. Gov't