Monoclonal antibody binding to human CD4 and its preparation method and application

By designing rabbit-derived recombinant antibodies conjugated with KLH using highly antigenic peptide fragments, the limitations of mouse-derived CD4 antibodies in terms of affinity and safety have been overcome, enabling efficient and highly specific detection of CD4-positive T cells and improving the detection accuracy and application scope of flow cytometry.

CN120623347BActive Publication Date: 2026-06-09JIANGSU ATAS BIOTECHNOLOGY CO LTD
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Patent Information

Application Number
CN202510802918.6
Authority / Receiving Office
CN · China
Patent Type
Patents(China)
Current Assignee / Owner
Filing Date
2025-06-16
Publication Date
2026-06-09
Estimated Expiration
2045-06-16

AI Technical Summary

Technical Problem

Existing murine CD4 antibodies have limitations in affinity, specificity and safety, and suffer from weak fluorescence signals and high background in flow cytometry, making it difficult to meet the increasingly demanding requirements for detection accuracy.

Method used

Rabbit recombinant antibody technology was used to design highly antigenic peptide fragments and conjugate them with KLH. Rabbits were then immunized with a rapid adjuvant, and high-affinity and highly specific rabbit recombinant antibodies were obtained through single B cell sorting and recombinant expression technology for the detection of CD4 positive T cells.

Benefits of technology

The obtained rabbit-derived recombinant antibody exhibited clear fluorescence signals and good background control in flow cytometry, significantly improving the sensitivity and accuracy of detection. It is applicable to fields such as flow cytometry, immunohistochemistry, magnetic bead sorting, HIV auxiliary diagnosis, and immune pathway research, and has broad application potential.

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Abstract

The application relates to the technical field of biological medicine, and particularly discloses a monoclonal antibody combined with human CD4 and a preparation method and application thereof. The antibody is obtained by combining single B cell sorting, gene cloning and mammalian cell expression technology after rabbit immunization of a CD4 polypeptide antigen. The antibody has high affinity and good specificity, and can effectively recognize CD4 positive T cells in human peripheral blood. The antibody is verified by ELISA and flow cytometry, and has high titer, low background and a signal-to-noise ratio superior to that of existing mouse-derived antibodies. Further, the amino acid sequences of the light chain and the heavy chain variable region of the antibody are obtained, so that controllability and repeatability of antibody expression are realized. The antibody can be widely applied to scenes such as immunodetection, HIV auxiliary diagnosis, T cell subgroup research and antibody engineering development, and has important scientific research and industrial values.
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