title: Human antibodies from phage libraries: neutralizing activity against human immunodeficiency virus type 1 equally improved after expression as Fab and IgG in mammalian cells.
abstract: Human antibodies against HIV-1 have been sought to study neutralization events on the molecular level, and for possible use in passive immune intervention. The development of phage display techniques has opened the possibility of rapidly generating human monoclonal antibodies with desired specificities. We and others have isolated human HIV-1 neutralizing antibody fragments using this technique. Bacterial expression of isolated clones does, however, differ broadly both in expression levels and functional activity. In addition, intact IgG cannot be expressed in bacteria. By transferring the genes of isolated Fab clones to a mammalian expression system we could perform a comparison of functional activity between Fab expressed in bacterial and mammalian cells, as well as Fab and whole IgG. Fab fragments expressed in mammalian cells showed increased virus neutralizing activity compared to the same Fab clones expressed in Escherichia coli, underlining the inefficiency of procaryotic expression. No difference in HIV-1 neutralizing capacity was detected between monovalent (Fab) and divalent (whole antibody) reagents expressed in CHO cells. Thus, bivalency does not always confer improved neutralization efficacy.
authors.affiliation: Karolinska Institute, Department of Medicine, Stockholm, Sweden.
authors.forename: A
authors.initials: A
authors.lastname: Samuelsson
authors.forename: F
authors.initials: F
authors.lastname: Yari
authors.forename: J
authors.initials: J
authors.lastname: Hinkula
authors.forename: O
authors.initials: O
authors.lastname: Ersoy
authors.forename: E
authors.initials: E
authors.lastname: Norrby
authors.forename: M A
authors.initials: MA
authors.lastname: Persson
mesh_terms: D000818:Animals; D000918:Antibody Specificity; D001667:Binding, Competitive / Q000276:immunology; D016466:CHO Cells; D003090:Coliphages / Q000276:immunology*; D006224:Cricetinae; D015723:Gene Library; D005822:Genetic Vectors / Q000276:immunology; D015483:HIV Antibodies / Q000096:biosynthesis* / Q000276:immunology*; D015497:HIV-1 / Q000276:immunology*; D006801:Humans; D006825:Hybridomas / Q000378:metabolism; D007140:Immunoglobulin Fab Fragments / Q000096:biosynthesis*; D007074:Immunoglobulin G / Q000096:biosynthesis*; D008969:Molecular Sequence Data; D009500:Neutralization Tests
chemical_list: D015483:HIV Antibodies; D007140:Immunoglobulin Fab Fragments; D007074:Immunoglobulin G