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CN-114829401-B - Anti-VHH domain antibodies and uses thereof

CN114829401BCN 114829401 BCN114829401 BCN 114829401BCN-114829401-B

Abstract

The present invention provides a set of anti-VHH domain antibodies and uses thereof. The invention further provides application of the antibody in development, screening and purification of nano-antibodies and application of the antibody in the field of immunotherapy.

Inventors

  • QIN XIJIAN
  • SUN LIWEI
  • WANG WANYI
  • SONG GUANGWEI

Assignees

  • 南京金斯瑞生物科技有限公司

Dates

Publication Date
20260505
Application Date
20200927
Priority Date
20190927

Claims (20)

  1. 1. An antibody or antigen binding fragment thereof that specifically binds to the VHH domain of a camelid antibody, wherein the antibody comprises a heavy chain variable region comprising HCDR1 shown as SEQ ID No. 14, HCDR2 shown as SEQ ID No. 29 and HCDR3 shown as SEQ ID No. 44 and a light chain variable region comprising LCDR1 shown as SEQ ID No. 59, LCDR2 shown as SEQ ID No. 74 and LCDR3 shown as SEQ ID No. 89.
  2. 2. The antibody or antigen-binding fragment thereof of claim 1, wherein the amino acid sequence of the heavy chain variable region is the amino acid sequence shown in SEQ ID No. 104 or an amino acid sequence having at least 90% identity thereto.
  3. 3. The antibody or antigen-binding fragment thereof of claim 1, wherein the amino acid sequence of the light chain variable region is the amino acid sequence set forth in SEQ ID No. 119 or an amino acid sequence having at least 90% identity thereto.
  4. 4. The antibody or antigen-binding fragment thereof of claim 1, which comprises the heavy chain variable region of SEQ ID No. 104 and the light chain variable region of SEQ ID No. 119.
  5. 5. The antibody or antigen-binding fragment thereof of claim 1, which specifically binds to a VHH domain with a binding affinity of K D of 10 nM to 1 pM.
  6. 6. The antibody or antigen binding fragment thereof of claim 1, wherein the camelid antibody is a single domain antibody or heavy chain antibody of dromedary (Camelus dromedarius), alpaca (Camelus bactrianus), alpaca (Vicugna pacos) or alpaca (LAMA GLAMA) origin.
  7. 7. The antibody or antigen binding fragment thereof of claim 1, which specifically binds to the VHH domain shown in SEQ ID No. 241 or a VHH domain having at least 60% amino acid sequence identity to SEQ ID No. 241.
  8. 8. The antibody or antigen binding fragment thereof of claim 1, which binds at the framework region of a VHH domain.
  9. 9. The antibody or antigen binding fragment thereof of claim 1, which binds at a conformational epitope of a VHH domain.
  10. 10. The antibody or antigen binding fragment thereof of claim 1, which binds at a conformational epitope of the framework region of a VHH domain.
  11. 11. The antibody or antigen binding fragment thereof of claim 1, which specifically binds to the framework region of the VHH domain shown in SEQ ID No. 241 or has at least 70% amino acid sequence identity to the framework region of the VHH domain shown in SEQ ID No. 241.
  12. 12. The antibody or antigen-binding fragment thereof of any one of claims 1 to 11, selected from the group consisting of Fab, F (ab') 2 , scFv, chimeric antibody, and humanized antibody.
  13. 13. One or more polynucleotides encoding the antibody or antigen-binding fragment thereof of any one of claims 1 to 12.
  14. 14. One or more vectors comprising the polynucleotide of claim 13.
  15. 15. The vector of claim 14, selected from the group consisting of cloning vectors and expression vectors.
  16. 16. A host cell comprising the polynucleotide of claim 13 or the vector of claim 14 or 15.
  17. 17. The host cell of claim 16, which is selected from a prokaryotic cell, a yeast cell, an insect cell, or a mammalian cell.
  18. 18. A method of producing an antibody or antigen-binding fragment thereof comprising culturing the host cell of claim 16 or 17 under conditions suitable for antibody production such that the antibody or antigen-binding fragment thereof is expressed.
  19. 19. The method of claim 18, further comprising recovering the antibody or antigen-binding fragment thereof.
  20. 20. A conjugate comprising the antibody or antigen-binding fragment thereof of any one of claims 1 to 12.

Description

Anti-VHH domain antibodies and uses thereof Technical Field The present invention relates to a set of anti-VHH domain antibodies. The invention also relates to a preparation method and an obtaining method of the antibody. Meanwhile, the invention relates to application of the antibody in development, screening and purification of nano antibodies. The invention also relates to the application of the antibody in the field of immunotherapy Background The camel-derived nano antibody (Nanobody) is also called as Single-domain antibody (Single-domain antibody) and heavy chain variable region antibody (variable domain of HEAVY CHAIN of HCAb, VHH), only comprises a variable region fragment of the camel-derived heavy chain antibody, has the size of about 15kDa, can be combined with antigen with high affinity and high specificity, and has very important application in the fields of antibody drug development, immune cell treatment and the like. In 1993 Hamers-Casterman et al found for the first time that the heavy chain antibody (heavy-chain antibodies,HCAbs)[C.Hamers,et al.,Naturally occurring antibodies devoid of light chains.Nature,1993.Vol 363:p 466-468],, which had a deletion of the light chain in the camelid, contained only the heavy chain variable region and the CH2 and CH3 constant regions. The VHH variable region expressed by molecular cloning means has very good stability and affinity and is the smallest antibody unit known to date. The Ablynx company has been successfully developed and marketed at present as a first therapeutic nanobody drug caplacizumab based on nanobody development technology for the treatment of acquired thrombotic thrombocytopenic purpura (aTTP) [ https:// www.ablynx.com/rd-portfolio/clinical-programmes/caplacizumab/]. Meanwhile, camel-derived nanobodies also have very important applications in the field of immune cell therapy. LCAR-B38M of Nanjing legend company is the first cell therapy reported clinically by cFDA in China and is also the first project for realizing the double report and double batch of Zhongmeishuangjia, which shows striking therapeutic effects in the data disclosed at present and has important significance in the development history of Chinese immunocytotherapy. The project adopts a unique nanometer antibody to design a chimeric antigen receptor on the design of the CAR, and avoids the defects of poor stability and low affinity of scFv in the conventional technical route. With the development of immune cell therapy technology, nanobodies are also becoming increasingly popular with researchers. Although nanobodies have important significance in the fields of antibody drug development and immune cell therapy, an antibody recognizing camel-derived nanobodies is currently lacking, so that the camel-derived nanobodies can be better developed or the identification, sorting and magnetic separation of CART cells in immune cell therapy can be optimized. The invention develops a group of high-affinity, high-specificity and high-functionality antibodies aiming at camel-source nano-antibodies, effectively solves the problems and meets the requirements of various application fields. Summary of The Invention In one aspect, the invention provides an antibody or antigen binding fragment thereof that specifically binds to a VHH domain. In one embodiment, the VHH domain is a VHH domain of a camelid antibody. In one embodiment, the camel-derived antibody is a single domain antibody or heavy chain antibody of dromedary (Camelus dromedarius), alpaca (Camelus bactrianus), alpaca (Vicugna pacos) or alpaca (LAMA GLAMA) origin. In another aspect, the invention provides an antibody or antigen-binding fragment thereof. In some embodiments, an antibody or antigen binding fragment thereof disclosed herein contains a Heavy Chain Variable Region (HCVR) and a Light Chain Variable Region (LCVR), wherein (a) the heavy chain variable region comprises heavy chain complementarity determining regions HCDR1, HCDR2, and HCDR3, (a) the HCDR1 sequence is selected from the group consisting of amino acid sequences shown in SEQ ID NOs 1,2, 3, 4, 5, 6,7, 8, 9, 10, 11, 12, 13, 14, or 15 or has at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, amino acid sequences thereof, An amino acid sequence of 99% or 100% identity, (b) the HCDR2 sequence is selected from the amino acid sequences shown in SEQ ID NOs 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29 or 30 or an amino acid sequence having at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% identity thereto, and (c) the HCDR3 sequence is selected from the group consisting of SEQ ID NOs 31, 32, 34, 35, 36, 37. 38, 39, 40, 41, 42, 43, 44, or 45, or an amino acid sequence having at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% identity thereto, (d) a heavy chain complementarity determining region HCDR comprising one or more amino acid sequences of (a), (B), and (c) deleted or inserted NO more than 3 amino acid substitut