Molecular cloning and primary characterization of a single-chain antibody against human sarcoma-associated antigen p200.
BACKGROUND
Monoclonal Antibody (MAb) 29-13 reacts with the human sarcoma-associated antigen (SAA) p200. We report here engineering and primary characterization of a single chain antibody (scFV2913).


METHODS
The scFV2913 recombinant gene, consisting of VH-linker-VK, was constructed with RT-PCR. This gene was cloned and expressed in E. coli. The renatured scFV2913 was used in the immunostaining study.


RESULTS
Consistent with its parent MAb 29-13, purified and renatured scFV2913 showed affinity and specificity to the SAA p200 according to the immuno-histochemical staining study of 99 specimens of human sarcomas and other tissues.


CONCLUSIONS
Due to its retained specificity and affinity, scFV2913 may be useful in immunodiagnosis and immunotherapy for sarcoma patients.
Chen
H
H
Department of Surgical Oncology, University of Illinois College of Medicine at Chicago, USA.
Wang
P
P
Stastny
J J
JJ
Shilkaitis
A
A
Liao
X
X
Das Gupta
T K
TK
D000595:Amino Acid Sequence; D000906:Antibodies / Q000235:genetics* / Q000276:immunology; D000911:Antibodies, Monoclonal / Q000235:genetics* / Q000276:immunology; D000951:Antigens, Neoplasm / Q000276:immunology*; D001483:Base Sequence; D003001:Cloning, Molecular; D006801:Humans; D007150:Immunohistochemistry; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction; D011994:Recombinant Proteins; D012509:Sarcoma / Q000276:immunology*; D057127:Single-Chain Antibodies
D000906:Antibodies; D000911:Antibodies, Monoclonal; D000951:Antigens, Neoplasm; D011994:Recombinant Proteins; D057127:Single-Chain Antibodies; C100570:antigen p200; C104655:recombinant antibody scFV2913