Immunological characterization of the human immunodeficiency virus type 1 reverse transcriptase protein by the use of monoclonal antibodies.
Eighteen monoclonal antibodies (MAbs) directed against the purified human immunodeficiency virus type 1 (HIV-1) reverse transcriptase (RT) protein were produced. The antibodies were characterized by competitive ELISAs and Western blot experiments, and with nested, nine amino acid long peptides representing the whole 560 amino acid RT protein. By ELISA, the MAbs react with a minimum of seven epitopes of the protein. Four of the epitopes were located on the N-terminal 51K subunit and the remaining three epitopes were located at the C-terminal end of the protein. Using synthetic peptides, two epitopes at the N-terminal part were located at amino acids 294 to 302 and 350 to 354, respectively, from the N-terminal start of the protein. One epitope was located at amino acids 442 to 450, just after the cleavage site between the N-terminal and C-terminal subunit at position 440. Antibodies located at amino acids 294 to 302 could inhibit the RT enzymic activity of the protein. Two other MAbs, directed at the N-terminal and C-terminal parts of the protein, could also inhibit RT activity.
Orvell
C
C
Department of Virology, National Bacteriological Laboratory, Stockholm, Sweden.
Unge
T
T
Bhikhabhai
R
R
Bäckbro
K
K
Rudén
U
U
Strandberg
B
B
Wahren
B
B
Fenyö
E M
EM
D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology*; D000956:Antigens, Viral / Q000276:immunology; D001667:Binding, Competitive; D015153:Blotting, Western; D004797:Enzyme-Linked Immunosorbent Assay; D000939:Epitopes; D005455:Fluorescent Antibody Technique; D015497:HIV-1 / Q000201:enzymology* / Q000276:immunology; D051379:Mice; D008807:Mice, Inbred BALB C; D012194:RNA-Directed DNA Polymerase / Q000276:immunology*
D000911:Antibodies, Monoclonal; D000956:Antigens, Viral; D000939:Epitopes; D012194:RNA-Directed DNA Polymerase