Polymerase chain reaction facilitates the cloning, CDR-grafting, and rapid expression of a murine monoclonal antibody directed against the CD18 component of leukocyte integrins.

Nucleic acids research ·Vol. 19 ·No. 9 ·1991-05-11 ·Pages 2471-6

Daugherty BL, DeMartino JA, Law MF, Kawka DW, Singer II, Mark GE

Abstract

Two novel approaches of recombinant PCR technology were employed to graft the complementarity determining regions from a murine monoclonal antibody (mAb) onto human antibody frameworks. One approach relied on the availability of cloned human variable region templates, whereas the other strategy was dependent only on human variable region protein sequence data. The transient expression of recombinant humanized antibody was driven by the adenovirus major late promoter and was detected 48 hrs post-transfection into non-lymphoid mammalian cells. The application of these new approaches enables the expression of a recombinant humanized antibody just 6 weeks after initiating the cDNA cloning of the murine mAb.

MeSH Terms

Animals Antibodies, Monoclonal/genetics,immunology Antigens, CD/immunology Base Sequence Binding Sites, Antibody/genetics CD18 Antigens Cells, Cultured Cloning, Molecular/methods DNA Gene Expression Genes, Immunoglobulin Genetic Vectors Humans Immunoglobulin Heavy Chains/genetics Immunoglobulin Light Chains/genetics Integrins/immunology Leukocytes/immunology Mice Molecular Sequence Data Polymerase Chain Reaction Recombinant Fusion Proteins/genetics,immunology Transfection

Chemicals

Antibodies, Monoclonal Antigens, CD CD18 Antigens Immunoglobulin Heavy Chains Immunoglobulin Light Chains Integrins Recombinant Fusion Proteins DNA

Authors & Affiliations

6 authors, click to expand affiliations / ORCID

Daugherty B L

Department of Cellular and Molecular Biology, Merck Sharp and Dohme Research Laboratories, Rahway, NJ 07065.

DeMartino J A

Law M F

Kawka D W

Singer I I

Mark G E

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Article Info

Journal

Nucleic acids research

Abbr.

Nucleic Acids Res

ISSN

0305-1048

DOI

10.1093/nar/19.9.2471

Published

1991-05-11

Pages

2471-6

Language

English

Region

England

NLM ID

0411011

PMCID

PMC329459

Subset

External Links

PubMed source DOI: 10.1093/nar/19.9.2471 PMC Full Text View journal details

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Polymerase chain reaction facilitates the cloning, CDR-grafting, and rapid expression of a murine monoclonal antibody directed against the CD18 component of leukocyte integrins.

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