TY - JOUR
T1 - Humanised recombinant antibody fragments bind human pancreatic islet cells
AU - Buenafe, Abigail C.
AU - Streeter, Philip
N1 - Funding Information:
Funding for this work was provided by the Juvenile Diabetes Research Foundation (grant no. 17-2013-327 to PRS).
Funding Information:
Funding for this work was provided by the Juvenile Diabetes Research Foundation (grant no. 17-2013-327 to PRS).
Publisher Copyright:
© 2018 Elsevier B.V.
PY - 2018/8
Y1 - 2018/8
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
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U2 - 10.1016/j.jim.2018.05.006
DO - 10.1016/j.jim.2018.05.006
M3 - Article
C2 - 29758224
AN - SCOPUS:85047270945
VL - 459
SP - 20
EP - 28
JO - Journal of Immunological Methods
JF - Journal of Immunological Methods
SN - 0022-1759
ER -