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Gelatin based substrate for protein-biochips

A protein and gelatin technology, applied in the field of protein microarray production, can solve the problems of complex and laborious surface modification methods, taking a long time, not large-scale industrial production, etc.

Inactive Publication Date: 2006-08-30
CARESTREAM HEALTH INC
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, all of these methods suffer from various disadvantages, either because of the long time required to prepare the surface or because the surface modification method is complex and laborious, making these methods not ideal for large-scale industrial production.

Method used

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  • Gelatin based substrate for protein-biochips
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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0076] This example illustrates the formulation of sandwich melts and gelatin overlayer melts and the method of coating the melts onto glass supports. This example illustrates the effectiveness of using an interlayer to provide the desired degree of adhesion of gelatin to a glass surface.

[0077] Recipe 1-1 (gelatin melt):

[0078] Solution 1: by adding 726.54 grams of type IV swelling gelatin (24.8% w / v) to 2237.06 grams of water, and adding 16 grams of nonylphenoxy polyglycerol coating aid and 20.4 grams of octylphenol poly(oxyethyl ) Sodium sulfonate coating assistant prepared.

[0079] Solution 2: Prepared by adding 800.79 grams of 1,1'-bis(vinyl(sulfonyl))methane (1.8% w / v) to 2199 grams of distilled water.

[0080] Prior to coating, equal volumes of solution 1 and solution 2 were mixed to form a single melt.

[0081] Recipe 1-2 (Interlayer Melt):

[0082] Prepared by adding 2.5 grams of gelatin, 16.3 grams of chromium potassium sulfate (chrom-alum), 34.7 grams of me...

Embodiment 2

[0087] This example illustrates the evaluation of gelatin-coated protein microarray substrates using a modified enzyme-linked immunosorbent assay (ELISA).

[0088] The operating procedure of the improved ELISA is as follows:

[0089] 1. Dissolve the goat anti-mouse antibody IgG from Sigma in PBS (phosphate-buffered saline, pH 7.4) buffer at a concentration of 1 mg / mL. A series of diluted goat anti-mouse IgG was hand-blotted onto nitrocellulose membranes and coated gelatin matrices. The dotted matrix was incubated for 1 hour at room temperature in a humidified cabinet.

[0090] 2. With 1% Triton X100 TM Wash the matrix four times with PBS buffer for 5 min each by shaking.

[0091] 3. The washed matrix was incubated in PBS buffer containing 1% glycine for 15 minutes with constant shaking.

[0092] 4. With 1% Triton TM Wash the matrix four times by shaking X100 in PBS buffer.

[0093] 5. Dilute Sigma's mouse IgG in a solution containing 0.1% Tween TM 20 in PBS buffer to 1...

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Abstract

A protein microarray element comprising: a) a support; b) a gelatin layer containing functional groups capable of binding biological probes; and interposed between the support and the gelatin layer and c) an adhesive interlayer layer capable of maintaining contact with the support and with the gelatin layer.

Description

field of invention [0001] The present invention relates generally to the fabrication of protein microarrays, and more particularly to methods employing gelatin-based matrices in which the gelatin surface has been modified to improve the specific attachment of biomolecules. Background of the invention [0002] The completion of the Human Genome Project stimulated the rapid development of the new interdisciplinary field of proteomics, which includes: the identification and characterization of the complete set of proteins encoded by the genome, protein synthesis, post-translational modifications, and detailed understanding of protein interactions at the level of cellular regulation. drawing. [0003] Although the combination of two-dimensional gel electrophoresis and mass spectrometry is still the main technology of proteomics research, the successful transplantation and application of DNA microarray technology in gene distribution and gene discovery has ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/543B01J19/00C07K17/00C40B40/10
CPCC07K17/00B01J2219/00659B01J2219/00641B01J2219/0074B01J2219/00725C40B40/10B01J19/0046
Inventor T·A·乔T·L·彭纳H·W·仇G·W·罗思
Owner CARESTREAM HEALTH INC
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