Abstract
A limiting dilution method for the efficient transformation by Epstein-Barr virus (EBV) of human B lymphocytes has been applied to the production of human monoclonal antibodies to ovarian cancer-associated antigens. Limited numbers (e.g., 2 × 105) of EBV-infected B lymphocytes from ovarian cancer patient spleen, lymph node, tumor, ascites and blood were successfully transformed using this method. An immunofiltration assay system was employed to identify EBV transformants secreting IgM antibody which reacted selectively with ovarian cancer patient ascites tumor cells, but not with a mixture of normal cell types. A miniature Western blot assay was utilized to screen for IgG reactivity to protein species in detergent extracts of ovarian cancer tumor cells. EBV-transformed cells selected after screening were then fused with heteromyeloma fusion partner SHM-D33 resulting in efficient recovery of hybridomas secreting MAb of the desired specificity. Human MAbs which selectively react with antigens associated with ovarian cancer tumor cells were obtained.
| Original language | English (US) |
|---|---|
| Pages (from-to) | 263-273 |
| Number of pages | 11 |
| Journal | Journal of Immunological Methods |
| Volume | 105 |
| Issue number | 2 |
| DOIs | |
| State | Published - Dec 24 1987 |
| Externally published | Yes |
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Keywords
- Epstein-Barr virus
- Monoclonal antibody, human
- Ovarian cancer-associated antigen
ASJC Scopus subject areas
- Biotechnology
- Immunology
Cite this
Generation of human monoclonal antibodies to cancer-associated antigens using limited numbers of patient lymphocytes. / Smith, Lloyd H; Yin, A.; Bieber, M.; Teng, N. N H.
In: Journal of Immunological Methods, Vol. 105, No. 2, 24.12.1987, p. 263-273.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - Generation of human monoclonal antibodies to cancer-associated antigens using limited numbers of patient lymphocytes
AU - Smith, Lloyd H
AU - Yin, A.
AU - Bieber, M.
AU - Teng, N. N H
PY - 1987/12/24
Y1 - 1987/12/24
N2 - A limiting dilution method for the efficient transformation by Epstein-Barr virus (EBV) of human B lymphocytes has been applied to the production of human monoclonal antibodies to ovarian cancer-associated antigens. Limited numbers (e.g., 2 × 105) of EBV-infected B lymphocytes from ovarian cancer patient spleen, lymph node, tumor, ascites and blood were successfully transformed using this method. An immunofiltration assay system was employed to identify EBV transformants secreting IgM antibody which reacted selectively with ovarian cancer patient ascites tumor cells, but not with a mixture of normal cell types. A miniature Western blot assay was utilized to screen for IgG reactivity to protein species in detergent extracts of ovarian cancer tumor cells. EBV-transformed cells selected after screening were then fused with heteromyeloma fusion partner SHM-D33 resulting in efficient recovery of hybridomas secreting MAb of the desired specificity. Human MAbs which selectively react with antigens associated with ovarian cancer tumor cells were obtained.
AB - A limiting dilution method for the efficient transformation by Epstein-Barr virus (EBV) of human B lymphocytes has been applied to the production of human monoclonal antibodies to ovarian cancer-associated antigens. Limited numbers (e.g., 2 × 105) of EBV-infected B lymphocytes from ovarian cancer patient spleen, lymph node, tumor, ascites and blood were successfully transformed using this method. An immunofiltration assay system was employed to identify EBV transformants secreting IgM antibody which reacted selectively with ovarian cancer patient ascites tumor cells, but not with a mixture of normal cell types. A miniature Western blot assay was utilized to screen for IgG reactivity to protein species in detergent extracts of ovarian cancer tumor cells. EBV-transformed cells selected after screening were then fused with heteromyeloma fusion partner SHM-D33 resulting in efficient recovery of hybridomas secreting MAb of the desired specificity. Human MAbs which selectively react with antigens associated with ovarian cancer tumor cells were obtained.
KW - Epstein-Barr virus
KW - Monoclonal antibody, human
KW - Ovarian cancer-associated antigen
UR - http://www.scopus.com/inward/record.url?scp=0023546921&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=0023546921&partnerID=8YFLogxK
U2 - 10.1016/0022-1759(87)90274-2
DO - 10.1016/0022-1759(87)90274-2
M3 - Article
C2 - 2826600
AN - SCOPUS:0023546921
VL - 105
SP - 263
EP - 273
JO - Journal of Immunological Methods
JF - Journal of Immunological Methods
SN - 0022-1759
IS - 2
ER -