Carbohydrate binding module and use thereof

一种糖类结合模块、聚糖结构的技术，应用在抑制HIV活性领域，能够解决未被实现、活性衰减、棘手安全问题等问题

Inactive Publication Date: 2013-05-08
张大慈
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, attenuation of the activity of HIV-related simian immunodeficiency virus, eliciting a protective response without resulting pathogenicity has not been achieved, raising safety concerns that make human experiments difficult

Method used

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  • Carbohydrate binding module and use thereof
  • Carbohydrate binding module and use thereof
  • Carbohydrate binding module and use thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0033] Structural relationship between SBD and 2G12

[0034] Structure-Based Multiple Sequence Alignment

[0035] Secondary structure prediction using Jpred server (server) (Cuff, J.A., Clamp, M.E., Siddiqui, A.S., Finlay, M. & Barton, GJ. JPred: a consensus secondary structure prediction server. Bioinformatics 14, 892-893 (1998)) and the Network Protein Sequence Analysis (NPSA) server. Jpred and NPSA servers provide consistent results from six (NNSSP, DSC, PREDATOR, MULPRED, ZPRED, and PHD) and twelve (SOPM, SOPMA, HNN, MLRC, DPM, DSC, GORI, GORII, GORIV, PHD , PREDATOR and SIMPA96) different methods.

[0036] result

[0037] Detection of structure-function relationships between RoSBD or AnSBD and other protein domains was performed by using the DALI database in silico (Holm, L., Kaariainen, S., Rosenstrom, P. & Schenkel, A. Searching protein structure databases with DaliLite v. 3. Bioinformatics 24, 2780-2781 (2008)). In total, 555 and 535 structurally related proteins ...

Embodiment 2

[0040] Functional relationship between SBD and 2G12

[0041] Microbes and Plasmids

[0042] Escherichia coli Top10F' (Invitrogen) was used for plasmid treatment, and E. coli BL21-Gold (DE3) (Invitrogen) was used for protein expression. Vectors pET-23a(+) and pET-15b (Novagen), both containing T7 primers (promoter), were used for expression of recombinant RoSBD and AnSBD in E. coli cells, respectively. Forward primer 5' for pET23a(+)-RoSBD

[0043] -CATATGGCAAGTATTCCCTAGCAGT-3' (SEQ ID No. 1) and reverse primer 5'-CTCGAGTCATGTAGATACTTGGT-3' (SEQ ID No. 2) containing NdeI and XhoI restriction sites, respectively, and forward primer for pET15b-AnSBD 5'-CATATGAGCAAGACCAGCACCAGT-3' (SEQ ID No. 3) and reverse primer 5'-CTCGAGCTACCGCCAGGTGT-3' (SEQ ID No. 4) containing NdeI and XhoI restriction sites, respectively, were used for cloning and sequence analysis.

[0044] Transformation and expression of SBD

[0045] The pET expression vector was transformed into Escherichia coli (E....

Embodiment 3

[0061] Specificity between RoSBD and AnSBD and HIV-1 glycoprotein

[0062] Competitive ELISA of HIV mAb and Glycans

[0063] The following peptides and mAbs were used in competitive ELISA experiments: (i) Human IgG 1 Secondary antibody ([2C11]; for human IgG 1 subclass-specific and not isotype-restricted; Fc-region-specific) secondary antibody, (ii) HIV-1 p24 peptide ([N29], a synthetic peptide based on N-terminal residues 1-104 of HIV-1 p24 ), (iii) HIV-1 gp120 peptide ([ED8.D4]; synthetic peptide of residues 427-448 of sequestered gp160 according to human HIV-1 BH8), (iv) HIV-1 gp41 peptide ([AG10H9], according to Synthetic peptide of residues 721-744 of HIV-1 gp160), (v) 2G12 mAb (recombinant human mAb against HIV-1 gp120), (vi) gp140 (HIV clade A, strain 92 / UG / 037). All were from Abeam, except 2G12 and gp140, which were purchased from Polymun Scientific.

[0064] For ELISA experiments, 50 μL of HIV-PC was mixed with an equal volume of 500 nM of each of (i) to (vi) abov...

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PUM

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Abstract

The present invention provides a method of inhibiting HIV infection of a subject, comprising administering to the subject an effective amount of an antibody mimetic of carbohydrate binding module (CBM) which specifically binds to an epitope on HIV glycoprotein. The present invention also provides a pharmaceutical composition comprising a therapeutically or prophylactically effective amount of an antibody mimetic of carbohydrate binding module and a pharmaceutically acceptable carrier. The present invention also provides a method of inhibiting HIV infection of the subject preventionally or therapically comprising administering to the subject an effective amount of an antibody mimetic of carbohydrate binding module (CBM) which specifically binds to an epitope on HIV glycoprotein

Description

field of invention [0001] The present invention relates to antibody mimetics of carbohydrate binding modules (CBMs) that specifically bind to epitopes of the HIV glycoprotein. The invention also relates to methods of inhibiting HIV activity. Background of the invention [0002] The human immunodeficiency virus (HIV) is known to cause acquired immunodeficiency syndrome (AIDS), and because HIV exhibits rapid genetic drift, widely divergent strains have emerged. Therefore, detection and treatment of variant strains has proven challenging and difficult. [0003] There is also an urgent need to develop effective prophylactic vaccines and other therapeutic strategies to limit HIV transmission while the epidemic continues unabated. The most successful vaccines consist of activity-attenuated or inactivated virus particles. However, attenuation of the activity of HIV-related simian immunodeficiency virus, eliciting a protective response without resulting pathogenicity has not been...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K16/08C12Q1/70
CPCC07K16/1063C07K2318/00C07K16/1045A61P31/18
Inventor 张大慈李远川黄荣渊林淑娟周维宜刘锡辉
Owner 张大慈
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