title: Recombinant human antibody fragment against tetanus toxoid produced by phage display.
abstract: Phage display technology is a powerful in vitro method for the identification of specific monoclonal antibodies (antibody fragments) to an antigenic target and allows the rapid generation and selection of high affinity, fully human antibodies directed toward any disease target appropriate for antibody therapy. In the present study, we exploited the phage display technology for the selection of an antigen binding fragment (Fabs) toward tetanus toxoid using human naïve phage antibody library constructed from peripheral blood lymphocytes of naïve human donors. The phages displaying Fab were subjected to three rounds of bio-panning with tetanus toxoid as antigen on a solid phase. The high affinity antibody fragments were expressed in HB2151 strain of Escherichia coli and purified by immobilized metal affinity chromatography. The binding activity and specificity of the antibody fragment was established by its reactivity toward tetanus toxoid and non-reactivity toward other related toxins as determined by enzyme-linked immunosorbent assay and immunoblot analysis. The selected Fab fragment forming the antigen-binding complexes with the toxoid in flocculation assay indicates that the Fab may have a potential neutralizing ability toward antigen.
authors.forename: B
authors.initials: B
authors.lastname: Neelakantam
authors.forename: N V
authors.initials: NV
authors.lastname: Sridevi
authors.forename: A M
authors.initials: AM
authors.lastname: Shukra
authors.forename: P
authors.initials: P
authors.lastname: Sugumar
authors.forename: S
authors.initials: S
authors.lastname: Samuel
authors.forename: L
authors.initials: L
authors.lastname: Rajendra
keywords: ELISA; Fabs; TT; flocculation; phage