Humanised recombinant antibody fragments bind human pancreatic islet cells

Abigail C. Buenafe, Philip Streeter

Research output: Contribution to journalArticle

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 languageEnglish (US)
JournalJournal of Immunological Methods
DOIs
StateAccepted/In press - Jan 1 2018

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Antibodies, Monoclonal, Humanized
Immunoglobulin Fragments
Islets of Langerhans
Endocrine Cells
Flow Cytometry
Immunohistochemistry
Monoclonal Antibodies
Hybridomas
Immune System
Enzyme-Linked Immunosorbent Assay
Antigens
Antibodies

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 journalArticle

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