Nanobody screening using sequence features

By utilizing the unique immune system of camels and next-generation sequencing technology, nanobodies with specific sequence characteristics were screened from a B cell library, solving the problem of insufficient epitope coverage in existing antibody technologies and achieving efficient and economical generation of nanobodies covering a wide range of antigenic epitopes.

JP2026521095APending Publication Date: 2026-06-26ZHEJIANG NANOMAB TECH CENT CO LTD +1
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Patent Information

Authority / Receiving Office
JP · JP
Patent Type
Applications
Current Assignee / Owner
ZHEJIANG NANOMAB TECH CENT CO LTD
Filing Date
2023-11-02
Publication Date
2026-06-26

AI Technical Summary

Technical Problem

Existing antibody technologies are unable to effectively cover the functional epitopes of target antigens, and the methods for generating antibodies are random, sporadic, and experimental, resulting in insufficient epitope coverage, selection redundancy, and low success rates.

Method used

By employing the unique immune system of camels and combining it with next-generation sequencing technology, nanobodies with specific sequence characteristics are screened from a library of camel B cells. Using specific amino acid positions and structural features, and through computer-aided selection and experimental verification, nanobodies covering a wide range of antigenic epitopes are generated.

Benefits of technology

This technology enables the large-scale, targeted generation of nanobodies covering a wide range of antigenic epitopes, improving selection efficiency and success rate while reducing costs.

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Abstract

A method for selecting camelid nanobodies from an array library collected from B cells of a camelid animal immunized with an antigen is provided. The method comprises (a) (i)phenylalanine (F) at position 42 (IMGT number), and (ii)a short hinge, and (iii)two or more cysteines in the nanobody sequence, and (iv)glutamine (Q) at position 123 (IMGT number), and (v)a low immunogenicity index, and (vi)a non-conventional VHH derived from germline IGHV3, or valine (V) included at position 42 (IMGT number), and (vii)a non-conventional VHH derived from germline IGHV4, or isoleucine (I) included at position 42 (IMGT number), and (viii)histidine (H), aspartic acid (D), or glutamic acid (E) in the CDR region, and (ix)histidine (H), aspartic acid (D), or glutamic acid (E) in the top 3 amino acid residues of the nanobody sequence, the FR2 region, or the top 16 amino acid residues of the FR3 region, and (x)tyrosine (Y) at position 42 (IMGT number), and a nanobody having a cyclic concave paratope structure arrangement, or (xi)phenylalanine (F) at position 42 (IMGT number), and a nanobody having a convex paratope structure arrangement, identifying a camelid nanobody having at least one of the features, and (b)measuring one or more biological activities of the nanobody identified in step (a).
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