Direct isolation of recombinant human antibodies against group B Neisseria meningitidis from scFv expression libraries.
The successful generation of human antibodies from large nai;ve antibody libraries requires iterative selection steps. Here, we describe a new and fast method for the isolation of high affinity antibodies directly from human single chain Fv antibody (scFv) expression libraries. Escherichia coli scFv expression libraries were made from peripheral blood lymphocytes from four individuals vaccinated with group B Neisseria meningitidis outer membrane vesicle (OMV) vaccine. Forty thousand clones were directly screened for antibodies binding N. meningitidis strain 44/76 (B:15:P1.7,16). Of 430 specific clones detected, 225 candidates were isolated and re-screened against the N. meningitidis strains NZ-98/254 (B:4:P1.7b,4) giving 4% cross-reactive clones. Antibodies were further characterized by DNA sequencing, ELISA and surface plasmon resonance (SPR) analysis, showing broad V-gene diversity and nanomolar scFv affinities. Antibodies derived by this method may assist in the discovery and development of new vaccine antigens as well as therapeutic antibody agents for the treatment of meningococcal diseases.
Stacy
John Erik
JE
Affitech AS, Gaustadalleen 21, N-0349 Oslo, Norway.
Kausmally
Louise
L
Simonsen
Bjørg
B
Nordgard
Silje H
SH
Alsøe
Lene
L
Michaelsen
Terje E
TE
Brekke
Ole Henrik
OH
D000595:Amino Acid Sequence; D000907:Antibodies, Bacterial / Q000302:isolation & purification*; D003429:Cross Reactions; D006801:Humans; D007128:Immunoglobulin Fragments / Q000276:immunology*; D007135:Immunoglobulin Variable Region / Q000235:genetics; D008969:Molecular Sequence Data; D009345:Neisseria meningitidis / Q000276:immunology*; D019151:Peptide Library; D011994:Recombinant Proteins / Q000302:isolation & purification*; D020349:Surface Plasmon Resonance
D000907:Antibodies, Bacterial; D007128:Immunoglobulin Fragments; D007135:Immunoglobulin Variable Region; D019151:Peptide Library; D011994:Recombinant Proteins; C055966:immunoglobulin Fv