Looking for breakthrough ideas for innovation challenges? Try Patsnap Eureka!

Nav1.7 antibodies and methods of using the same

A nav1.7, antibody technology, applied in chemical instruments and methods, antibodies, anti-receptor/cell surface antigen/cell surface determinant immunoglobulin, etc., can solve problems such as poor display selectivity and side effects

Inactive Publication Date: 2016-07-06
DUKE UNIV
View PDF155 Cites 0 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, these drugs at 9 Na V Shows poor selectivity among channel subtypes and, therefore, nonselectively targets multiple Na V Channel isoforms can lead to off-target effects, which in turn lead to serious side effects (e.g. cardiotoxicity)

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
  • Nav1.7 antibodies and methods of using the same
  • Nav1.7 antibodies and methods of using the same
  • Nav1.7 antibodies and methods of using the same

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 2-12

[0549] Materials and methods for Examples 2-12

[0550] Antibody production and purification. Mouse monoclonal antibodies were generated by Abmart using peptides having the following sequences: HHPMTEEFKN (115 (also referred to herein as CTmab); SEQ ID NO: 20) and VELFLADVEG (1E16 (also referred to herein as SVmab1); SEQ ID NO: 21). Both antibodies are IgG1. Upon receipt of the hybridomas, multiple rounds of limiting dilution cloning are performed to select for stable monoclonal cell populations. Using recombinant Na V 1.7 Enzyme-linked immunoassay (ELISA) assay of DIIVSD was used for screening. The hybridoma cells of 1E16 and 1I5 were incubated in a hollow fiber bioreactor (Fibercell Systems Inc, US), and then the supernatant was collected every two days. Supernatants from each harvest were screened by ELISA assay and purified on protein G sepharose columns following the manufacturer's protocol (Invitrogen, US).

[0551] Whole-cell patch-clamp recordings of HEK293 cells....

Embodiment 2

[0574] Monoclonal Antibody (mAb) Production

[0575] According to bacterial Na V Channel Na V For the crystal structure of Ab, the choice is equivalent to Na V1.7 The peptide of the tip (ring) of the DII voltage sensor switch (i.e., the S3-S4 loop) instead of using the complete channel as the antigen for antibody production ( Figure 4 B). Also choose the equivalent of Na V The peptide of the DIIS1-S2 loop of 1.7 was used as a negative control, because the S1-S2 loop does not move when the membrane potential changes, so the mAb binding to the S1-S2 loop will not significantly affect the Na V 1.7 channel gating ( Figure 4 C). One mAb was raised against each of the following regions: 1E16 mAb (i.e. sodium channel voltage sensor mAb (hence also referred to herein as SVmab1)) recognizes Na V The S3-S4 loop of 1.7, and the control 1I5 mAb (also referred to herein as CTmab) recognize Na V 1.7 S1-S2 ring. Both antibodies belonged to the same subtype and showed positive ELIS...

Embodiment 3

[0577] 1E16mAb stabilizes Na in an application (state)-dependent manner V 1.7's closed state

[0578] To test the 1E16 and 1I5mAbs against Na V 1.7 Effect of Transiently Expressed Na Using a Whole-Cell Voltage Clamp Configuration V 1.7 HEK293 cells were subjected to electrophysiological recording (ie, patch clamp recording). Current traces were elicited by 30 ms voltage steps between −80 and +60 mV in 10 mV increments from a membrane clamping potential of −120 mV. When 1 μM 1I5 mAb was added to the extracellular side, no significant change was observed in the peak sodium current ( Image 6 A and 6B). However, 100 nM 1E16 mAb produced a significant reduction in peak sodium current ( Image 6 C and 6D). Comparison of the conductance-voltage relationship of the control 1I5mAb- and 1E16mAb-improved currents when 100 nM 1E16mAb was added showed a shift in depolarization (approximately 20 mV) ( Image 6 E). In contrast, comparison of the steady-state inactivation curves show...

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
electrical resistanceaaaaaaaaaa
Login to View More

Abstract

Disclosed herein is an antibody that binds to a voltage sensor paddle (VSP) of Nav1.7. Also disclosed herein are methods of treating pain, itch, neurogenic inflammation, or cough in a subject in need thereof. The methods include administrating the antibody to the subject.

Description

[0001] Cross References to Related Applications [0002] This application claims U.S. Provisional Patent Application No. 61 / 874,234, filed September 5, 2013, U.S. Provisional Patent Application No. 61 / 915,304, filed December 12, 2013, and U.S. Provisional Patent Application No. 61 / 915,304, filed December 12, 2014 61 / 944,388, which is hereby incorporated by reference in its entirety. [0003] Statement of Government Rights [0004] This invention was made with Government support under Contract Nos. NIH1DP2OD008380-01, R01DE17794, R01DE22743, and R01NS67686 awarded by the National Institutes of Health. The US Government has certain rights in this invention. technical field [0005] The present invention relates to anti-Na V 1.7 Antibodies and use of said antibodies to detect and / or inhibit Na V 1.7 method. Antibodies can be used to treat a subject with disease, cough, pain and / or itching. Background of the invention [0006] Voltage-gated sodium (Na V ) channel (Voltage...

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
Patent Type & Authority Applications(China)
IPC IPC(8): C07K16/28
CPCA61K2039/505A61P11/06A61P17/04A61P17/06A61P25/04A61P25/06C07K16/28C07K2299/00C07K2317/33C07K2317/34C07K2317/76C07K2317/92A61K39/395C07K14/705C07K2317/55C07K2317/56
Inventor 李硕容纪如荣
Owner DUKE UNIV
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Patsnap Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Patsnap Eureka Blog
Learn More
PatSnap group products

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

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap|About US| Contact US: help@patsnap.com