Human single chain monoclonal antibody that recognizes matrix protein of heterologous influenza A virus subtypes.
Matrix protein (M1) is predominant and has pivotal role in the influenza A virus replication and assembly. It is therefore an attractive target for antiviral drugs, siRNA studies, and therapeutic antibodies. Nevertheless, therapeutic antibody that interferes with the M1 multiplex function has never been developed. In this study, human single monoclonal antibody fragments (HuScFvs) to M1 were generated. Full length recombinant M1 (rM1) was produced from cDNA prepared from genome of highly pathogenic avian influenza virus, A/H5N1. The rM1 was used as an antigen in phage bio-panning to select phage clones displaying HuScFv from a human antibody phage display library. Several phage clones displaying HuScFv bound to the rM1 and harboring the respective huscfv gene inserts were isolated. RFLP experiments revealed multiple DNA banding patterns which indicated epitope/affinity diversity of the HuScFv. The HuScFv were tested for their binding to native M1 of homologous and heterologous influenza A viruses using ELISA as well as incorporating immunostaining and immunofluorescence studies with infected MDCK cells. One such protein produced from a selected phage clone blocked binding of M1 to viral RNA. The HuScFv in their in vivo functional format, e.g. cell-penetrating molecules, should be developed and tested as a broad spectrum anti-A/influenza.
Poungpair
Ornnuthchar
O
Graduate Program, Faculty of Allied Health Science, Thammasat University, Pathumthani 12120, Thailand.
Chaicumpa
Wanpen
W
Kulkeaw
Kasem
K
Maneewatch
Santi
S
Thueng-in
Kanyarat
K
Srimanote
Potjanee
P
Tongtawe
Pongsri
P
Songserm
Thaweesak
T
Lekcharoensuk
Porntippa
P
Tapchaisri
Pramuan
P
D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology*; D000914:Antibodies, Viral / Q000276:immunology; D000918:Antibody Specificity; D004285:Dogs; D004797:Enzyme-Linked Immunosorbent Assay; D006801:Humans; D050440:Immunoglobulin Subunits / Q000276:immunology; D017404:In Situ Hybridization, Fluorescence; D009980:Influenza A virus / Q000276:immunology* / Q000378:metabolism; D051379:Mice; D019151:Peptide Library; D011817:Rabbits; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000276:immunology; D014763:Viral Matrix Proteins / Q000096:biosynthesis / Q000276:immunology*
D000911:Antibodies, Monoclonal; D000914:Antibodies, Viral; D050440:Immunoglobulin Subunits; C488936:M1 protein, Influenza A virus; D019151:Peptide Library; D011994:Recombinant Proteins; D014763:Viral Matrix Proteins