Investigation of interactions between binding residues and solubility of grafted humanized anti-VEGF IgG antibodies expressed as full-length format in the cytoplasm of a novel engineeredE. coliSHuffle strain

บทความในวารสาร

ผู้เขียน/บรรณาธิการ

กลุ่มสาขาการวิจัยเชิงกลยุทธ์

รายละเอียดสำหรับงานพิมพ์

รายชื่อผู้แต่ง: Sangpheak K., Waraho-Zhmayev D., Haonoo K., Torpaiboon S., Teacharsripaiboon T., Rungrotmongkol T., Poo-Arporn R.P.

ผู้เผยแพร่: Royal Society of Chemistry

ปีที่เผยแพร่ (ค.ศ.): 2021

วารสาร: RSC Advances (2046-2069)

Volume number: 11

Issue number: 11

หน้าแรก: 6035

หน้าสุดท้าย: 6048

จำนวนหน้า: 14

นอก: 2046-2069

eISSN: 2046-2069

URL: https://www.scopus.com/inward/record.uri?eid=2-s2.0-85100810246&doi=10.1039%2fd0ra08534k&partnerID=40&md5=11b0be48bbe52eef9ef73c25b8e3f7ae

ภาษา: English-Great Britain (EN-GB)

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บทคัดย่อ

Monoclonal antibodies (mAbs) are one of the fastest-growing areas of biopharmaceutical industry and have been widely used for a broad spectrum of diseases. Meanwhile, the immunogenicity of non-human derived antibodies can generate side effects by inducing the human immune response to produce human anti-mouse-immunoglobulin antibody (HAMA). In this work, we aim to reduce the immunogenicity of muMAb A.4.6.1 by substitute human sequences for murine sequences. Humanized antibodies are constructed by grafting, specificity determining residues (SDR), complementary determining regions (CDR), and chimeric region of muMAb A.4.6.1, onto variable domain of Trastuzumab (Herceptin). The interactions between grafted antibodies and their target, Vascular endothelial growth factor (VEGF), were theoretically investigated by molecular dynamics simulation in order to evaluate the antibodies-antigen binding behavior. The obtained protein-protein interactions and calculated binding free energy suggested that the SDR-VEGF complex presented a significantly greater binding affinity, number of contact and total number of H-bonds compared to CDR and chimeric mAbs, significantly. Moreover, the Camsol program predicted that the solubility of SDR mAb exhibits the greatest solubility. This result was supported by performing a western blot analysis of the grafted mAbs with soluble and insoluble fractions in order to evaluate their solubility, in which SDR was found to have a much lower amount of insoluble proteins. Consequently, the enhanced binding affinity and solubility of the designed SDR was achieved by the single S106D mutation using computational methods. With the aim of low immunogenicity, high solubility, and high affinity, this SDR humanized antibody was expected to have greater efficacy than murine or chimeric antibodies for future use in humans. © The Royal Society of Chemistry 2021.

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