# US 2014/0193840 A1 · Chromatography filter paper-based elisa

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- Number: US 2014/0193840 A1
- Document ID: US20140193840A1
- Status: Published application
- Kind: A1
- Application number: 13/867,409
- Filed: 2013-04-22
- Published: 2014-07-10
- Earliest claimed: 2013-01-08
- Assignees: National Tsing Hua University
- Applicants: National Tsing Hua University
- Inventors: Min-Yen HSU; Keng-Hung LIN; Chao-Min CHENG
- Web page: https://patentgrep.com/patent/US20140193840A1

## CPC

- G01N 33/548: Carbohydrates, e.g. dextran
- G01N: INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES (measuring or testing apparatus or processes other than immunoassay, involving enzymes or microorganisms C12M, C12Q)

## Abstract

A chromatography filter paper-based ELISA includes following steps: providing a chromatography filter paper plate provided with a plurality of independent hydrophilic regions defined by at least one hydrophobic region; immobilizing an antigen of a sample onto one of the hydrophilic regions of the chromatography filter paper plate; and detecting the antigen with an ELISA process, wherein an antibody used in the ELISA process is a monoclonal antibody. The present invention has advantages in lower amount of loaded sample, higher sensitivity and rapid analysis in comparison to conventional ELISA.

## Claims

### Claim 1

A chromatography filter paper-based ELISA comprising:
providing a chromatography filter paper plate having a plurality of independent hydrophilic regions defined by at least one hydrophobic region;
immobilizing an antigen of a sample onto one of the hydrophilic regions of the chromatography filter paper plate; and
detecting the antigen by an ELISA process, wherein a monoclonal antibody is used in the ELISA process.

### Claim 2 (depends on 1)

The chromatography filter paper-based ELISA according to claim 1, wherein the chromatography filter paper plate is made of cotton fibers.

### Claim 3 (depends on 1)

The chromatography filter paper-based ELISA according to claim 1, wherein the hydrophobic region is formed by wax printing.

### Claim 4 (depends on 1)

The chromatography filter paper-based ELISA according to claim 1, wherein the antigen is a growth factor.

### Claim 5 (depends on 1)

The chromatography filter paper-based ELISA according to claim 1, wherein the sample is aqueous humor or vitreous humor.

### Claim 6 (depends on 1)

The chromatography filter paper-based ELISA according to claim 1, wherein the antigen is an angiogenic factor.

### Claim 7 (depends on 6)

The chromatography filter paper-based ELISA according to claim 6, wherein the angiogenic factor comprises vascular endothelial growth factor (VEGF), fibroblast growth factor (FGF), platelet-derived endothelial growth factor (PDEGF) or angiopoietin.

### Claim 8 (depends on 5)

The chromatography filter paper-based ELISA according to claim 5, wherein the antigen is vascular endothelial growth factor (VEGF).

### Claim 9 (depends on 5)

The chromatography filter paper-based ELISA according to claim 5, wherein the monoclonal antibody is Bevacizumab or Ranibizumab.

### Claim 10 (depends on 1)

The chromatography filter paper-based ELISA according to claim 1, wherein the antigen is dispersed in a liposome or a microsphere.

### Claim 11 (depends on 1)

The chromatography filter paper-based ELISA according to claim 1 further comprising:
determining the amount of the monoclonal antibody so as to detect the concentration of the antigen.

### Claim 12 (depends on 11)

The chromatography filter paper-based ELISA according to claim 11, wherein a colorimetric enzyme conjugated to the monoclonal antibody is used to determine the amount of the monoclonal antibody via detecting a colorimetric reaction of the colorimetric enzyme.

### Claim 13 (depends on 11)

The chromatography filter paper-based ELISA according to claim 11, wherein the colorimetric enzyme is a horseradish peroxidase (HRP).

### Claim 14 (depends on 11)

The chromatography filter paper-based ELISA according to claim 11, wherein the colorimetric enzyme comprises alkaline phosphatase, beta-galactosidase, glucose oxidase or glucose-6-phosphate dehydrogenase.

### Claim 15 (depends on 11)

The chromatography filter paper-based ELISA according to claim 11 further comprising:
using a second antibody to bind to the monoclonal antibody to determine the amount of the monoclonal antibody via detecting the amount of the second antibody.