Construction Of Human Lymphatic Filariasis Antibody Phage Display Library And Production Of Recombinant Monoclonal Antibodies Against Bmr1 And Bmsxp Filarial Antigens
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Date
2018-01
Authors
Rahumatullah, Anizah
Journal Title
Journal ISSN
Volume Title
Publisher
Universiti Sains Malaysia
Abstract
A Global Programme for Elimination of Lymphatic Filariasis aims to eliminate the disease as a public health problem by the year 2020. Brugia Rapid and PanLF Rapid are rapid diagnostic tools that are being used in this programme. They are based on the detection of recombinant proteins BmR1 and BmSXP. The quality of these tests should be well-maintained at all time or even improved as the GPELF progresses and a sensitive and specific test may be needed at the end of the programme and for surveillance post-certification. To address this need, the availability of recombinant monoclonal antibodies to the two recombinant proteins would be very useful. The first objective of this study involved the construction of a human lymphatic filariasis scFv library using phage display technology. The library was generated using a TA based intermediate shuttle system for cloning and the diversity of the library was 108. Then monoclonal antibodies were isolated via biopanning using the newly constructed immune library and an in-house produced naïve library. From the immune library six and two monoclonal antibodies to BmSXP and BmR1, respectively were isolated. From the naïve library two monoclonal antibodies to each recombinant protein were isolated. All the monoclonal antibodies were characterized based on their V-gene usage, gene pairing, CDR length, amino acid distribution, polarity and position. The results showed that the immune library has both kappa and lambda family gene representation, however only lambda family clones were isolated from the naïve library.
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Keywords
Construction of human lymphatic filariasis antibody , recombinant monoclonal antibodies against BmR1