Abstract
A novel cell display system was developed for cloning the variable region (V) genes of antigen-specific human antibodies. The system is based on an antibody library displayed on the surface of COS cells, using a plasmid vector designed to direct expression of membrane-bound antibodies. COS cells expressing antigen-specific antibodies were separated using a flow cytometer for their binding to a fluorescent dye-labeled antigen. To test the performance of this system, we cloned V genes of 4 antibodies directed against hepatitis B surface antigen (HBsAg) from a library prepared from peripheral blood lymphocytes of a vaccinated donor. These membrane-bound anti-HBsAg antibodies were easily converted to soluble forms, all of which showed a size similar to human serum IgG in SDS-PAGE and the same specific binding to HBsAg as membrane-bound forms in ELISA. All V(H) and Y(κ) gene segments of the 4 clones isolated in this study belonged to V(H)III and V(κ)I subgroups, respectively. These findings demonstrate the potential and selection capabilities of our cell display system for cloning the V genes of antigen-specific human antibodies.
| Original language | English |
|---|---|
| Pages (from-to) | 193-204 |
| Number of pages | 12 |
| Journal | Journal of Immunological Methods |
| Volume | 202 |
| Issue number | 2 |
| DOIs | |
| Publication status | Published - 28-03-1997 |
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All Science Journal Classification (ASJC) codes
- Immunology and Allergy
- Immunology
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Cell display library for gene cloning of variable regions of human antibodies to hepatitis B surface antigen. / Higuchi, Kazuo; Araki, Takeyoshi; Matsuzaki, Osamu; Sato, Akiko; Kanno, Kimiyoshi; Kitaguchi, Nobuya; Ito, Hirataka.
In: Journal of Immunological Methods, Vol. 202, No. 2, 28.03.1997, p. 193-204.Research output: Contribution to journal › Article
TY - JOUR
T1 - Cell display library for gene cloning of variable regions of human antibodies to hepatitis B surface antigen
AU - Higuchi, Kazuo
AU - Araki, Takeyoshi
AU - Matsuzaki, Osamu
AU - Sato, Akiko
AU - Kanno, Kimiyoshi
AU - Kitaguchi, Nobuya
AU - Ito, Hirataka
PY - 1997/3/28
Y1 - 1997/3/28
N2 - A novel cell display system was developed for cloning the variable region (V) genes of antigen-specific human antibodies. The system is based on an antibody library displayed on the surface of COS cells, using a plasmid vector designed to direct expression of membrane-bound antibodies. COS cells expressing antigen-specific antibodies were separated using a flow cytometer for their binding to a fluorescent dye-labeled antigen. To test the performance of this system, we cloned V genes of 4 antibodies directed against hepatitis B surface antigen (HBsAg) from a library prepared from peripheral blood lymphocytes of a vaccinated donor. These membrane-bound anti-HBsAg antibodies were easily converted to soluble forms, all of which showed a size similar to human serum IgG in SDS-PAGE and the same specific binding to HBsAg as membrane-bound forms in ELISA. All V(H) and Y(κ) gene segments of the 4 clones isolated in this study belonged to V(H)III and V(κ)I subgroups, respectively. These findings demonstrate the potential and selection capabilities of our cell display system for cloning the V genes of antigen-specific human antibodies.
AB - A novel cell display system was developed for cloning the variable region (V) genes of antigen-specific human antibodies. The system is based on an antibody library displayed on the surface of COS cells, using a plasmid vector designed to direct expression of membrane-bound antibodies. COS cells expressing antigen-specific antibodies were separated using a flow cytometer for their binding to a fluorescent dye-labeled antigen. To test the performance of this system, we cloned V genes of 4 antibodies directed against hepatitis B surface antigen (HBsAg) from a library prepared from peripheral blood lymphocytes of a vaccinated donor. These membrane-bound anti-HBsAg antibodies were easily converted to soluble forms, all of which showed a size similar to human serum IgG in SDS-PAGE and the same specific binding to HBsAg as membrane-bound forms in ELISA. All V(H) and Y(κ) gene segments of the 4 clones isolated in this study belonged to V(H)III and V(κ)I subgroups, respectively. These findings demonstrate the potential and selection capabilities of our cell display system for cloning the V genes of antigen-specific human antibodies.
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UR - http://www.scopus.com/inward/citedby.url?scp=0031588945&partnerID=8YFLogxK
U2 - 10.1016/S0022-1759(97)00010-0
DO - 10.1016/S0022-1759(97)00010-0
M3 - Article
VL - 202
SP - 193
EP - 204
JO - Journal of Immunological Methods
JF - Journal of Immunological Methods
SN - 0022-1759
IS - 2
ER -