Abstract
We describe here the humanisation of two mouse monoclonal antibodies that bind to surface markers on human pancreatic islet endocrine cells. Monoclonal antibodies produced by the HIC1-2B4 and HIC0-4F9 mouse hybridomas bind distinct surface molecules expressed on endocrine cells and have been validated for a number of experimental methods including immunohistochemistry and live cell sorting by flow cytometry. Variable region framework and first constant region domain sequences were replaced with that from compatible human antibody sequences, and the resulting recombinant antigen-binding fragments were cloned and expressed in mouse myeloma cells. ELISA, fluorescent immunohistochemistry, and flow cytometry were used to assess the specificity of the humanised antibody fragments. Purification and binding analyses indicated that human islet endocrine cell binding was retained in the humanised antibody fragments. These humanised, recombinant antibody fragments have a lower risk of eliciting adverse responses from a patient's immune system and, therefore, have highly improved clinical potential. Thus, they may be used to image, target or carry cargo specifically to islet cells in human patients.
| Original language | English (US) |
|---|---|
| Journal | Journal of Immunological Methods |
| DOIs | |
| State | Accepted/In press - Jan 1 2018 |
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Keywords
- Beta cells
- Humanisation
- Islets
- Monoclonal antibody
- Recombinant antibody, antigen-binding fragment
ASJC Scopus subject areas
- Immunology and Allergy
- Immunology
Cite this
Humanised recombinant antibody fragments bind human pancreatic islet cells. / Buenafe, Abigail C.; Streeter, Philip.
In: Journal of Immunological Methods, 01.01.2018.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - Humanised recombinant antibody fragments bind human pancreatic islet cells
AU - Buenafe, Abigail C.
AU - Streeter, Philip
PY - 2018/1/1
Y1 - 2018/1/1
N2 - We describe here the humanisation of two mouse monoclonal antibodies that bind to surface markers on human pancreatic islet endocrine cells. Monoclonal antibodies produced by the HIC1-2B4 and HIC0-4F9 mouse hybridomas bind distinct surface molecules expressed on endocrine cells and have been validated for a number of experimental methods including immunohistochemistry and live cell sorting by flow cytometry. Variable region framework and first constant region domain sequences were replaced with that from compatible human antibody sequences, and the resulting recombinant antigen-binding fragments were cloned and expressed in mouse myeloma cells. ELISA, fluorescent immunohistochemistry, and flow cytometry were used to assess the specificity of the humanised antibody fragments. Purification and binding analyses indicated that human islet endocrine cell binding was retained in the humanised antibody fragments. These humanised, recombinant antibody fragments have a lower risk of eliciting adverse responses from a patient's immune system and, therefore, have highly improved clinical potential. Thus, they may be used to image, target or carry cargo specifically to islet cells in human patients.
AB - We describe here the humanisation of two mouse monoclonal antibodies that bind to surface markers on human pancreatic islet endocrine cells. Monoclonal antibodies produced by the HIC1-2B4 and HIC0-4F9 mouse hybridomas bind distinct surface molecules expressed on endocrine cells and have been validated for a number of experimental methods including immunohistochemistry and live cell sorting by flow cytometry. Variable region framework and first constant region domain sequences were replaced with that from compatible human antibody sequences, and the resulting recombinant antigen-binding fragments were cloned and expressed in mouse myeloma cells. ELISA, fluorescent immunohistochemistry, and flow cytometry were used to assess the specificity of the humanised antibody fragments. Purification and binding analyses indicated that human islet endocrine cell binding was retained in the humanised antibody fragments. These humanised, recombinant antibody fragments have a lower risk of eliciting adverse responses from a patient's immune system and, therefore, have highly improved clinical potential. Thus, they may be used to image, target or carry cargo specifically to islet cells in human patients.
KW - Beta cells
KW - Humanisation
KW - Islets
KW - Monoclonal antibody
KW - Recombinant antibody, antigen-binding fragment
UR - http://www.scopus.com/inward/record.url?scp=85047270945&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=85047270945&partnerID=8YFLogxK
U2 - 10.1016/j.jim.2018.05.006
DO - 10.1016/j.jim.2018.05.006
M3 - Article
AN - SCOPUS:85047270945
JO - Journal of Immunological Methods
JF - Journal of Immunological Methods
SN - 0022-1759
ER -