Expression of recombinant human antibody fragments capable of inhibiting the phospholipase and myotoxic activities of Bothrops jararacussu venom.
Phospholipases A(2) are components of Bothrops venoms responsible for disruption of cell membrane integrity via hydrolysis of its phospholipids. This study used a large nonimmune human scFv library named Griffin.1 (MRC, Cambridge, UK) for selection of recombinant antibodies against antigens present in Bothrops jararacussu venom and identification of specific antibodies able to inhibit phospholipase activity. Four clones were identified as capable of inhibiting this activity in vitro. These clones were able to reduce in vivo the myotoxic activity of BthTX-I and BthTX-II PLA(2), but had no effect on the in vitro anticoagulant activity of BthTX-II. This work shows the potential of using recombinant scFv libraries in the search for antibodies that neutralize relevant venom components.
Tamarozzi
M B
MB
Programa de Pós-Graduação em Imunologia Básica e Aplicada, Faculdade de Medicina de Ribeirão Preto, Brazil.
Soares
S G
SG
Marcussi
S
S
Giglio
J R
JR
Barbosa
J E
JE
D000818:Animals; D000906:Antibodies / Q000235:genetics / Q000276:immunology* / Q000302:isolation & purification / Q000378:metabolism*; D001777:Blood Coagulation / Q000276:immunology; D017837:Bothrops / Q000378:metabolism*; D003435:Crotalid Venoms / Q000201:enzymology* / Q000276:immunology / Q000633:toxicity*; D004797:Enzyme-Linked Immunosorbent Assay; D015870:Gene Expression; D054501:Group II Phospholipases A2; D006321:Heart / Q000187:drug effects*; D006801:Humans; D007700:Kinetics; D051379:Mice; D010446:Peptide Fragments / Q000235:genetics / Q000276:immunology / Q000302:isolation & purification / Q000378:metabolism; D010741:Phospholipases A / Q000276:immunology / Q000378:metabolism / Q000633:toxicity*; D011994:Recombinant Proteins / Q000235:genetics / Q000276:immunology / Q000302:isolation & purification / Q000378:metabolism; D030162:Reptilian Proteins; D012995:Solubility
D000906:Antibodies; D003435:Crotalid Venoms; D010446:Peptide Fragments; D011994:Recombinant Proteins; D030162:Reptilian Proteins; C078097:bothropstoxin; D010741:Phospholipases A; D054501:Group II Phospholipases A2; C072031:bothropstoxin II