Identification of human antibody fragment clones specific for tetanus toxoid in a bacteriophage. lambda. immunoexpression library

Journal Article · Mon Oct 01 04:00:00 EDT 1990 · Proceedings of the National Academy of Sciences of the United States of America; (USA)

DOI:https://doi.org/10.1073/pnas.87.20.8095· OSTI ID:5803655

Mullinax, R L; Gross, E A; Amberg, J R; Hogrefe, H H; Kubitz, M M; Greener, A; Alting-Mees, M; Ardourel, D; Short, J M; Sorge, J A ^[1]; Hay, B N; Shopes, B ^[2]

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1. Stratagene, La Jolla, CA (USA)
2. Stratacyte, La Jolla, CA (USA)

The authors have applied a molecular biology approach to the identification of human monoclonal antibodies. Human peripheral blood lymphocyte mRNA was converted to cDNA and a select subset was amplified by the polymerase chain reaction. These products, containing coding sequences for numerous immunoglobulin heavy- and {kappa} light-chain variable and constant region domains, were inserted into modified bacteriophase {lambda} expression vectors and introduced into Escherichia coli by infection to yield a combinatorial immunoexpression library. Clones with binding activity to tetanus toxoid were identified by filter hybridization with radiolabeled antigen and appeared at a frequency of 0.2{percent} in the library. These human antigen binding fragments, consisting of a heavy-chain fragment covalently linked to a light chain, displayed high affinity of binding to tetanus toxoid with equilibrium constants in the nanomolar range but did not cross-react with other proteins tested. They estimate that this human immunoexpression library contains 20,000 clones with high affinity and specificity to our chosen antigen.

• OSTI ID: 5803655
• Journal Information: Proceedings of the National Academy of Sciences of the United States of America; (USA), Journal Name: Proceedings of the National Academy of Sciences of the United States of America; (USA) Vol. 87:20; ISSN 0027-8424; ISSN PNASA
• Country of Publication: United States
• Language: English

Related Subjects

550201* -- Biochemistry-- Tracer Techniques
59 BASIC BIOLOGICAL SCIENCES
ANTIBODIES
BACTERIAL DISEASES
BACTERIOPHAGES
BETA DECAY RADIOISOTOPES
DAYS LIVING RADIOISOTOPES
DISEASES
DNA HYBRIDIZATION
DNA POLYMERASES
DNA SEQUENCING
ELECTRON CAPTURE RADIOISOTOPES
ENZYMES
GENE AMPLIFICATION
HYBRIDIZATION
IMMUNOTHERAPY
INFECTIOUS DISEASES
INTERMEDIATE MASS NUCLEI
IODINE 125
IODINE ISOTOPES
ISOTOPES
MICROORGANISMS
MOLECULAR BIOLOGY
MONOCLONAL ANTIBODIES
NUCLEI
NUCLEOTIDYLTRANSFERASES
ODD-EVEN NUCLEI
PARASITES
PHOSPHORUS-GROUP TRANSFERASES
POLYMERASES
RADIOISOTOPES
STRUCTURAL CHEMICAL ANALYSIS
TETANUS
THERAPY
TOXOIDS
TRANSFERASES
VIRUSES