Human neutralizing human immunodeficiency virus type 2-specific Fab molecules generated by phage display.
A panel of human immunodeficiency virus type 2 (HIV-2)-neutralizing, recombinant Fab fragments was generated by using the phage display technique. The combinatorial library was derived from an asymptomatic, HIV-2-seropositive individual and constructed on the surface of filamentous phage by using the pComb3 phagemid vector and then screened against native HIV-2 envelope glycoprotein (gp125). Ten of 30 Fab fragments generated displayed strong reactivity in an ELISA and were therefore selected for further study. Six of these possessed neutralizing capacity, with titres varying from 20 to 80 against the homologous HIV-2 strain, and one also had a weak neutralizing capacity against a heterologous HIV-2 isolate, K135. Sequencing of the heavy chain CDR3 regions showed that the gp125-specific Fabs represented individual clones. These reagents may be useful for studies on the conformational structures of the HIV-2 envelope antigens and their immunogenicity, which may help in vaccine design. Furthermore, the cloned Fab genes may be transformed into whole IgG for eukaryotic expression, and as such used for therapeutic and immunoprophylactic studies in HIV-2-infected macaques and, possibly, for human immunoprophylaxis against HIV-2.
Björling
Ewa
E
Department of Immunology, Microbiology, Pathology and Infectious Diseases (IMPI), Karolinska Institute, Huddinge, Sweden2.|Microbiology and Tumorbiology Center (MTC), Karolinska Institute, S-171 77 Stockholm, Sweden1.
von Garrelts
Eva
E
Department of Medicine, Karolinska Institute, Karolinska Hospital (L8:01), S-171 76 Stockholm, Sweden3.
Mörner
Andreas
A
Department of Immunology, Microbiology, Pathology and Infectious Diseases (IMPI), Karolinska Institute, Huddinge, Sweden2.|Microbiology and Tumorbiology Center (MTC), Karolinska Institute, S-171 77 Stockholm, Sweden1.
Ehnlund
Mariethe
M
Microbiology and Tumorbiology Center (MTC), Karolinska Institute, S-171 77 Stockholm, Sweden1.
Persson
Mats A A
MAA
Department of Medicine, Karolinska Institute, Karolinska Hospital (L8:01), S-171 76 Stockholm, Sweden3.
D000595:Amino Acid Sequence; D000918:Antibody Specificity; D001435:Bacteriophages; D004797:Enzyme-Linked Immunosorbent Assay; D015686:Gene Products, env / Q000276:immunology*; D015483:HIV Antibodies / Q000737:chemistry / Q000276:immunology*; D015488:HIV Antigens / Q000276:immunology*; D015658:HIV Infections / Q000097:blood / Q000821:virology; D015498:HIV-2 / Q000276:immunology*; D006801:Humans; D007140:Immunoglobulin Fab Fragments / Q000737:chemistry / Q000276:immunology*; D007143:Immunoglobulin Heavy Chains / Q000276:immunology; D008297:Male; D008875:Middle Aged; D008969:Molecular Sequence Data; D009500:Neutralization Tests; D019151:Peptide Library; D011498:Protein Precursors / Q000276:immunology*; D054299:env Gene Products, Human Immunodeficiency Virus
D015686:Gene Products, env; D015483:HIV Antibodies; D015488:HIV Antigens; D007140:Immunoglobulin Fab Fragments; D007143:Immunoglobulin Heavy Chains; D019151:Peptide Library; D011498:Protein Precursors; D054299:env Gene Products, Human Immunodeficiency Virus; C066316:gp125 envelope protein, Human immunodeficiency virus 2