A human monoclonal antibody scFv to urokinase plasminogen activator.
A human monoclonal antibody can be a good method for tumor diagnosis and treatment. This study is aimed at the generation of human antibody fragments against urokinase plasminogen activator (uPA) known to be related to tumor metastasis using the naive human antibody phage display library. Three clones--A2, A8, and E4--were selected from 1 x 10(10) sized human naïve antibody phage library using BIAcore rescue and screen. Clone A8 was finally selected by flow cytometry against Hep3 and HT1080, uPA overexpressing tumor cell lines. A8 clone consisted of 324 bp lambda and 402 bp heavy chains. The affinity (K(D)) of purified A8 antibody fragments was 1.44 x 10(-8) M(-1). The antibody fragment was reacted with HT1080 in a dose-dependent manner but not reacted with LS513 normal fibroblast. In this study, uPA specific human monoclonal antibody fragment A8 was made with BIAcore selection. Selected A8 was bound specifically to uPA expressed on the tumor cell surface. Further study for the application of A8 antibody clones will be needed.
Bae
Ki-Beom
KB
Department of Surgery, Pusan Paik Hospital, Busan, Republic of Korea.
Jeong
Young-Joo
YJ
Won
Hae Jeong
HJ
Hong
Kwan-Hee
KH
Choi
Il-Whan
IW
Seo
Su-Kil
SK
Park
Sae-Gwang
SG
D000595:Amino Acid Sequence; D000911:Antibodies, Monoclonal / Q000096:biosynthesis* / Q000235:genetics / Q000276:immunology*; D000915:Antibody Affinity; D001483:Base Sequence; D045744:Cell Line, Tumor; D017931:DNA Primers / Q000235:genetics; D005434:Flow Cytometry; D006801:Humans; D008969:Molecular Sequence Data; D017422:Sequence Analysis, DNA; D057127:Single-Chain Antibodies / Q000235:genetics / Q000276:immunology*; D020349:Surface Plasmon Resonance; D014568:Urokinase-Type Plasminogen Activator / Q000276:immunology*
D000911:Antibodies, Monoclonal; D017931:DNA Primers; D057127:Single-Chain Antibodies; D014568:Urokinase-Type Plasminogen Activator