Unlock instant, AI-driven research and patent intelligence for your innovation.

Solid-phase n-terminal peptide capture and release

A solid support and composition technology, applied in the preparation method of peptides, carrier binding/immobilizing peptides, peptides, etc., can solve harmful and other problems

Pending Publication Date: 2021-06-22
BOARD OF RGT THE UNIV OF TEXAS SYST
View PDF3 Cites 0 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Therefore, there is a limit to what can be obtained, and any loss of sample is extremely detrimental to proper analysis

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
  • Solid-phase n-terminal peptide capture and release
  • Solid-phase n-terminal peptide capture and release
  • Solid-phase n-terminal peptide capture and release

Examples

Experimental program
Comparison scheme
Effect test

preparation example Construction

[0297] Preparation of Immobilization Resin - Method A: Protide-amine polystyrene resin (CEM Corporation) was coupled with 3 different linkers: (1) Fmoc-Rink linker, (2) no linker, or (3) three glycine residues base. The linker was coupled with HCTU (4 equiv) and DIEA (8 equiv)) at 4.4 equiv for 20 minutes each. For these linkers, 6-formylpyridine-2-carboxylic acid (Enamine) (2.2 equiv) was coupled using HCTU (2 equiv) and diisopropylethylamine (6 equiv) in DMF for 1 hour at room temperature. It was then washed extensively with DMF and stored at 4 °C.

[0298] Aldehyde Trap Resin Preparation - Method B: Fmoc-Peg2-OH, Rink linker and 6-formylpyridine-2-carboxylate for chemical coupling using amino-PEGA resin (Novabiochem) and using HCTU / DIEA (1:1:1.1 ratio) Acid functionalization for 45 minutes. Deprotection was performed twice with 20% piperidine in DMF for five minutes each. The resin was stored in DMF at 4°C until use.

[0299] Peptide Capture: Remove the resin and allow...

example 2

[0321] Example 2 - Labeling of captured peptides

[0322] After the peptide has been captured by the peptide resin, any desired chemical reaction can be performed. This includes isobaric, fluorescent, biotin or PEG labeling of proteins as well as acetylation or other capping steps required prior to analysis. It also allows performing many of these steps in a manner similar to solid-phase peptide synthesis without subsequent purification steps ( Image 6 ).

example 3

[0323] Example 3 - Probe Design

[0324] The resin can be designed and synthesized to contain a linker between the capture moiety (eg, PCA) and the support. A unique identifier such as an oligomer (eg, DNA, RNA, PNA) or a tandem mass tag (TMT or TMT) can be incorporated onto the adapter or support. Examples of probe designs are depicted in Figures 7A and 7B. The probes in Figures 7A and 7B represent probes containing nucleic acid barcode sequences, but the nucleic acid barcode sequences can be replaced with barcodes as described herein.

[0325] Other such designs can be envisaged if cutting from beads is not required. For example, a probe may not contain a cleavable unit. The probe can be constructed with a cleavable group in the linker, and the peptide can be cleaved from the probe via the cleavable group. Then, depending on its use, the PCA adduct is removed by using a hydrazine type release agent. Thus, a two-step release process is possible. Even without performing ...

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
thicknessaaaaaaaaaa
Login to View More

Abstract

Provided herein are rapid and reversible methods to non-specifically immobilize peptides and proteins irrespective of their sequence, as well as small molecules, on a solid support to allow for manipulations of and reactions with these molecules in a manner that does not require purification between steps, which increases sample yield and reduces the quantity of starting material required.

Description

[0001] This application claims the benefit of priority to U.S. Provisional Application Serial No. 62 / 741,833, filed October 5, 2018, and U.S. Provisional Application Serial No. 62 / 879,735, filed July 29, 2019, the entire contents of both applications Incorporated herein by reference. [0002] Statement Regarding Federally Sponsored Research [0003] This invention was made with government support under Grant No. R35 GM122480 awarded by the National Institutes of Health. The government has certain rights in this invention. Background technique [0004] Chemical manipulation of proteins and peptides is a common method for attaching a variety of linkers, including isobaric moieties or isotope-labeled chemicals for comparative mass spectrometry studies (Weise et al., 2007), allowing quantitative measurements of binding constants (Andrews et al. et al., 2008), and installation of a purified shank (Klement et al., 2010). For these experiments, samples must be purified from remain...

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): C07K14/00
CPCC07K17/14C07K17/06B01D15/265B01J20/3204B01J20/3219B01J20/3274B01J20/3272B01J20/3285B01J20/3293B01J20/3248C12Q1/6804G01N33/54353G01N2458/10C12Q2525/161C12Q2563/179C12Q2565/514G01N33/68C07K1/042B01J20/0229B01J20/22G01N33/6824
Inventor E·安斯林E·马科特C·J·霍华德二世J·斯瓦米纳坦A·M·巴尔多J·鲁瑟B·M·弗洛伊德
Owner BOARD OF RGT THE UNIV OF TEXAS SYST
Features
  • R&D
  • Intellectual Property
  • Life Sciences
  • Materials
  • Tech Scout
Why Patsnap Eureka
  • Unparalleled Data Quality
  • Higher Quality Content
  • 60% Fewer Hallucinations
Social media
Patsnap Eureka Blog
Learn More

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

© 2025 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap|About US| Contact US: help@patsnap.com