Sustained-release anti-FcRn antibody or antigen binding fragment and application thereof

A technology for combining fragments and antibodies, which is applied in the field of slow-release anti-FcRn antibodies or antigen-binding fragments, treatment, prevention and/or diagnosis of FcRn-related diseases, and can solve the problem of short half-life of FcRn antibodies or Fc fragments, fast metabolism of antibody drugs, etc. question

Active Publication Date: 2022-01-11
BEIJING KOHNOOR SCI & TECH CO LTD
View PDF9 Cites 1 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] The present invention aims to solve the problem of fast metabolism of some antibody drugs in the p

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • Sustained-release anti-FcRn antibody or antigen binding fragment and application thereof
  • Sustained-release anti-FcRn antibody or antigen binding fragment and application thereof
  • Sustained-release anti-FcRn antibody or antigen binding fragment and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0043] Example 1: Nanobody construction

[0044] Camels were immunized with antigens, peripheral blood mononuclear cells (PBMC) were isolated and total RNA was extracted for reverse transcription, and the variable domain of the heavy-chain of heavy chain antibody was amplified using the reverse transcription product as a template , VHH) and connected to the phage display vector, electroporated into Escherichia coli TG1 competent cells, and camel immune library was constructed.

[0045] Specifically, camels are immunized once every two weeks, a total of 4 times. Each injection of 0.8 mg HSA extracellular region recombinant protein (purchased from Hualan Bioengineering Co., Ltd., product batch number: 201405030), accompanied by Freund's complete / incomplete adjuvant (Sigma, F5881, F5506), was taken subcutaneously at multiple points The way of injection. Two weeks after each immunization, 1 mL of blood was collected to separate the serum, and the immunogen was used as the assay ...

Embodiment 2

[0047] Example 2: Construction of FcRn sustained release antibody

[0048] In this example, the HSA nanobody HzH4-3 and Hz2MG6m19 sequences obtained in Example 1 were connected to the C-terminus of the FcRn antibody Efgartigimod (Ef for short, the sequence is shown in SEQ ID NO: 9) to construct H4-3-Ef and m19 -Ef eukaryotic expression vector, and then transfected into eukaryotic cells for expression and purification to obtain slow-release antibodies against hFcRn H4-3-Ef and m19-Ef.

Embodiment 3

[0049] Example 3: ELISA method to detect the metabolism of H4-3-Ef and m19-Ef in HSA / FcRn double-transformed humanized mice

[0050] The experiment was conducted with hFcRn female 6-week-old transgenic mice, divided into 3 groups, 4 mice in each group, and the first two groups were intravenously injected with H4-3-Ef and m19-Ef, both at 300 μg / mouse. Then blood was collected by docking at 2h, 4h, 8h, 24h, 48h, 79h, 120h, 151h and 192h. Another group was injected with Ef at a rate of 300 μg / bird, and then blood was collected at 0.5h, 1h, 3h, 4h, 5h, 6h, 8h, 24h, 30h, 48h, 79h and 96h, and the serum was collected and stored at -20°C. save.

[0051] After all the blood collection is completed, the collected serum is tested by ELISA, and the steps are as follows:

[0052] 1) Coating anti-Fc Ab (Sigma, I2136) onto 96-well ELISA plate, 0.5 μg / mL, 100 μL / well, overnight at 4°C;

[0053] 2) After washing the plate 3 times with PBS, add 0.5% casein-PBS, block at 37°C for 60 min, and...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to view more

PUM

PropertyMeasurementUnit
Absorbanceaaaaaaaaaa
Login to view more

Abstract

The invention discloses a sustained-release anti-FcRn antibody or antigen binding fragment and application thereof. The sustained-release anti-FcRn antibody or antigen binding fragment comprises a heavy chain variable region and a heavy chain constant region. The heavy chain variable region is VHH of a heavy chain antibody of camelidae or a polypeptide consisting of the VHH of the heavy chain antibody of the camelidae; the VHH comprises CDRs 1, 2 and 3; the VHH CDR1 region, the VHH CDR2 region and the VHH CDR3 region respectively comprise amino acid sequences shown as SEQ ID NO: 1, SEQ ID NO: 2 and SEQ ID NO: 3 or amino acid sequences with at least 80% identity with amino acid sequences shown as SEQ ID NO: 4, SEQ ID NO: 5 and SEQ ID NO: 6; and the heavy chain constant region has an amino acid sequence which is at least 80% identical with an amino acid sequence as shown in SEQ ID NO: 9. By conjugating an HSA antibody sequence to an FcRn antibody, the half-life period of an original antibody is prolonged, and the number of times of drug administration is reduced.

Description

technical field [0001] The invention relates to the field of biological medicine. Specifically, the present invention relates to a novel artificially designed antibody with extended half-life, especially a slow-release anti-FcRn antibody or antigen-binding fragment. The present invention also relates to the therapeutic and diagnostic uses of these FcRn-binding antibodies, especially in the treatment, prevention and / or diagnosis of FcRn-related diseases, such as autoimmune-related diseases. Background technique [0002] For the problem of fast metabolism of some antibodies, several methods have been developed in the past to prolong the half-life of antibody fragments and prepare sustained-release drugs. These methods include reducing the sensitivity of the fragment to serum proteases using specific slow-release formulations, or reducing the intrinsic clearance of antibody fragments by amino acid substitutions that reduce receptor binding affinity in the endosomal compartment...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to view more

Application Information

Patent Timeline
no application Login to view more
IPC IPC(8): C07K16/28C12N15/13A61K39/395A61K47/68A61P37/02
CPCC07K16/2833A61K47/6803A61K47/6849A61P37/02C07K2317/22C07K2317/52C07K2317/565C07K2317/569A61K2039/505
Inventor 王荣娟张畅曾大地焦莎莎王双张锦超
Owner BEIJING KOHNOOR SCI & TECH CO LTD
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Try Eureka
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap