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

Techniques for predicting, detecting and reducing aspecific protein interference in assays involving immunoglobulin single variable domains

A technology of immunoglobulin and structural domain, applied in the field of immunoglobulin single variable domain, which can solve the problem of not providing

Pending Publication Date: 2018-10-16
ABLYNX NV
View PDF27 Cites 0 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0016] However, none of these references are aware that certain proteins present in the subject's blood or serum would interfere with ADA assays involving ISV's, and because of this, these references do not address (nor provide protocols for) The question of how to avoid non-specific protein interference in said ADA assay, thereby allowing the ADA assay to be used to determine the true presence / extent of anti-drug antibodies (emerging or already present) in the sample to be tested

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
  • Techniques for predicting, detecting and reducing aspecific protein interference in assays involving immunoglobulin single variable domains
  • Techniques for predicting, detecting and reducing aspecific protein interference in assays involving immunoglobulin single variable domains
  • Techniques for predicting, detecting and reducing aspecific protein interference in assays involving immunoglobulin single variable domains

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0231] Example 1: Generation of Antibodies for Polyclonal Analysis.

[0232] Polyclonal antibodies (IgG fraction) that can be used as "analytical antibodies" are produced as follows:

[0233] a. Identification of suitable plasma samples for isolation of polyclonal antibodies

[0234] Twenty plasma samples from healthy individuals who had never been treated with ISV were evaluated for the presence of antibodies against ISV useful as analytical antibodies of the present invention.

[0235] The ISV used initially in this example was SEQ ID NO:1. Subsequently, the assay described below was repeated with other ISV's in order to confirm that the interaction was not specific to this particular ISV, but rather a non-specific protein-protein interaction that may occur with many ISV's (see paragraph C) below). As an alternative to SEQ ID NO: 1, eg SEQ ID NO: 2 can also be used.

[0236] The assay used was an ECL (electrochemiluminescence) based bridging assay using biotinylated ISV...

Embodiment 2

[0260] Example 2: Analysis of affinity purification of antibodies

[0261] This example describes two methods that can be used to isolate analytical antibodies capable of recognizing and / or binding to the C-terminal end of ISV from biological fluids from human subjects. Antibodies were isolated from 4 different serum samples characterized in that these induced a positive signal in the ADA assay tested according to Example 1.

[0262] Starting from a serum sample, each of these methods provides a purified preparation of an interfering factor that can be used as an analytical antibody in the methods described herein. These methods can also be used, more generally, to purify interfering factors for other purposes (for example, interfering factors purified using the protocol below were also used experimentally in Example 8 to show that ISV or ISV-constructs were associated with monoclonal 21-4 Binding of the same ISV or ISV-construct with interfering factors, and thus predicts ...

Embodiment 2A

[0263] Example 2A: Purification Using Protein A and Affinity Chromatography

[0264] In the first step, IgG antibody fractions are enriched from serum samples using protein A affinity chromatography. Typical columns used for this enrichment include HiTrap MabselectSure and MabSelectXtra (GE Healthcare); PorosMabCapture A (Applied Biosystems). In all experiments, IgG antibodies were purified from serum samples in an automated and analogous manner. Chromatographic protocols were performed on an AKTA purification system (GE Healthcare) and recorded in real time using UNICORN protein purification software (GE Healthcare). Briefly, serum samples were diluted 1:1 with D-PBS (Dulbecco's Phosphate Buffered Saline) and filtered at 0.22 μm before loading onto the column at a fixed flow rate of 0.5 mL / min. The column was washed with D-PBS at a flow rate of 0.5 mL / min to remove non-specifically bound components within 5 column volumes. The IgG fraction was eluted by acidic elution us...

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

No PUM Login to View More

Abstract

The invention provides, and in certain specific but non-limiting aspects, methods for modifying and / or improving ISV's so as to eliminate or reduce their propensity to produce such protein interference or signal; a modification of the ISV can be introduced that eliminates or reduces the propensity of the ISV to produce the protein interference or the signal; ISV's specifically selected (eg, usingthe assays described herein) such that it does not have or has a lower / reduced propensity for protein interference or the signal; modified and / or improved ISV's that do not have or have a lower / reduced propensity to produce the protein interference or the signal .

Description

[0001] This application is a Chinese patent application No. with an application date of June 25, 2012 and the title of the invention is "Technology for Predicting, Detecting and Reducing Non-specific Protein Interference in Assays Involving Immunoglobulin Single Variable Domains" .201280030902.9 divisional application. technical field [0002] The present invention relates to the field of immunoglobulin single variable domains. Background technique [0003] An immunoglobulin single variable domain, or "ISV," is generally defined herein as an amino acid sequence that [0004] -comprises an immunoglobulin fold or is capable of forming an immunoglobulin fold (i.e. by folding) under suitable conditions (such as physiological conditions), i.e. thereby forming an immunoglobulin variable domain (such as, for example, VH, VL or VHH domain); [0005] and [0006] -forms (or is capable of forming under such suitable conditions) an immunoglobulin variable domain comprising a functio...

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): C07K16/00A61K39/395
CPCA61K2039/505C07K16/00C07K16/2875C07K16/42C07K2317/22C07K2317/31C07K2317/35C07K2317/567C07K2317/569C07K2317/92G01N33/5306C07K2317/34G01N33/54393G01N33/6857A61P37/04C07K16/4283C07K2317/94G01N33/6854C07K16/28A61K39/39533C07K2319/30C07K16/18C07K2317/76
Inventor 尤迪特·鲍迈斯特玛丽-葆拉·吕西安娜·阿曼达·布什卡罗·布东玛丽-安格·比斯维尔勒·斯诺尔克斯特凡妮·斯塔埃朗
Owner ABLYNX NV
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