[A specific human scFv against malignant melanoma selected from a large human phage antibody library].
OBJECTIVE
To obtain the specific human scFv against malignant melanoma (MM) using phage antibody library technology.


METHODS
The library was panned with a certain kind of MM cells for 4 rounds. The antigen binding activities of random clones were tested by ELISA in order to select specific antibodies, which were then examined by DNA sequence analysis and immunohistochemistry.


RESULTS
In ELISA and immunohistochemical staining, the positive clone selected from the 80 random clones was able to bind one kind of MM cell specifically, while not able to bind other irrelavant cells or antigens, such as human's squamous carcinoma cells, keratinocytes, melanocytes, keratin, trypsinase, transferrin and mouse IgG, etc. The antibody's V(H) genes belonged to human IgG V(H)5 subgroup and the V(L) belonged to human V(kappa) subtype determined by DNA sequence analysis.


CONCLUSIONS
One specific human phage antibody against MM cells was obtained by phage display technique, which may be useful for further development of tumor-targeted therapies.
Li
Miao
M
Department of Dermatology Xijing Hospital, Fourth Military Medical University, Xi'an 710032, China. youlonglm@sina.com
Chen
Yu-ping
YP
Wang
Gang
G
Li
Chun-ying
CY
Yin
He-hui
HH
Tian
Yan-li
YL
Gao
Tian-wen
TW
D000906:Antibodies / Q000235:genetics* / Q000276:immunology*; D000918:Antibody Specificity; D004797:Enzyme-Linked Immunosorbent Assay; D006801:Humans; D007074:Immunoglobulin G / Q000235:genetics / Q000276:immunology; D007135:Immunoglobulin Variable Region / Q000235:genetics* / Q000276:immunology*; D007150:Immunohistochemistry; D008545:Melanoma / Q000276:immunology; D019151:Peptide Library; D017422:Sequence Analysis, DNA
D000906:Antibodies; D007074:Immunoglobulin G; D007135:Immunoglobulin Variable Region; D019151:Peptide Library