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Isolation and characterization of camelid single-domain antibodies against HER2.

ABSTRACT:

Objective

To isolate and characterize novel high-affinity llama single-domain antibodies against human HER2.

Results

We immunized a llama with human HER2, constructed a phage-displayed V_HH library from the lymphocytes of the animal, and isolated six unique HER2-specific V_HHs by panning. All six V_HHs were unique at the amino acid level and were clonally unrelated, as reflected by their distinct CDR3 lengths. All six V_HHs recognized recombinant human HER2 ectodomain with monovalent affinities ranging from 1 to 51 nM, had comparable affinities for cynomolgus monkey HER2, and bound HER2⁺ SKOV3 cells by flow cytometry. Three of the V_HHs recognized recombinant murine HER2 with no loss of affinity compared with human and cynomolgus monkey HER2. The V_HHs recognized three major epitopes on HER2 (including one conserved across the human, simian and murine orthologues), all of which were distinct from that of trastuzumab. These V_HHs may be useful in the design of modular cancer immunotherapeutics.

SUBMITTER: Hussack G

PROVIDER: S-EPMC6282393 | biostudies-literature | 2018 Dec

REPOSITORIES: biostudies-literature

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Isolation and characterization of camelid single-domain antibodies against HER2.

Hussack Greg G Raphael Shalini S Lowden Michael J MJ Henry Kevin A KA

BMC research notes 20181205 1

<h4>Objective</h4>To isolate and characterize novel high-affinity llama single-domain antibodies against human HER2.<h4>Results</h4>We immunized a llama with human HER2, constructed a phage-displayed V<sub>H</sub>H library from the lymphocytes of the animal, and isolated six unique HER2-specific V<sub>H</sub>Hs by panning. All six V<sub>H</sub>Hs were unique at the amino acid level and were clonally unrelated, as reflected by their distinct CDR3 lengths. All six V<sub>H</sub>Hs recognized recomb ...[more]

PMID: 30518413

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