Antigen detection kit and method

A technology for antigen detection and kits, which is applied in the field of antigen detection kits and methods, and can solve problems such as difficult operation

Inactive Publication Date: 2011-03-23
KOREA INST OF SCI & TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Although in principle this method is considered a multiplex ELISA, in practice it becomes more difficult to perform as the number of analytes to be analyzed in a sample increases beyond 2

Method used

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  • Antigen detection kit and method

Examples

Experimental program
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Effect test

Embodiment 1

[0062] Example 1: OLISA analysis using RNase H for a single antigen

[0063] Detection of alpha-fetoprotein (AFP, from human fetal umbilical cord serum, Fitzgerald, Inc.) was performed as follows.

[0064] 1.1 Preparation of detection probe (d-Ab-DNA)

[0065] 50 μl of 3.4 mg / mL monoclonal antibody against AFP (1) (Cat# 10C-CR1007M5, which recognizes AFP from cord blood) for sandwich ELISA provided in a pair by Fitzgerald, Inc. was mixed with 50 μl of PBS The buffer was mixed with 1 μl of 100 mM SMPH (succinimidyl-6-[β-maleimidopropionamido]hexanoate) and incubated at room temperature for 30 minutes. The obtained solution was diluted in 15 mL of PBS buffer, and centrifuged at 4,000 rpm at 4°C for 40 minutes with Amicon Ultra-15 (Ultracel-30K) (Millipore). The process was repeated 3 times, and the supernatant ① was obtained.

[0066] At the same time, add 100 μl of 100 μM DNA solution (the DNA nucleotide sequence is 5'-TTTTTTTTTTTTTTTTTTTTTAACCACAGTG-3', SEQ ID NO: 1, in whi...

Embodiment 2

[0072] Example 2: Multiplex-OLISA analysis using RNase H for simultaneous analysis of two types of antigens coexisting in the same sample

[0073]The following experiments for the simultaneous detection of alpha-fetoprotein (AFP) and prostate specific antigen (PSA, Fitzgerald, Cat# 30C-CP1017U) were performed.

[0074] 2.1 Preparation of detection probe (d-Ab-DNA)

[0075] AFP detection probes were prepared in the same manner as described in Example 1.1.

[0076] The PSA detection probe was prepared as follows. 50 µl of the antibody solution at a concentration of 2.25 mg / mL was mixed with 50 µl of PBS buffer and 1 µl of 100 mM SMPH (succinimidyl-6-[β-maleimidopropionamido]hexanoate). The antibody solution contains monoclonal antibodies against PSA (3), Cat#10-P20D, supplied by Fitzgerald Inc. in paired form for total PSA analysis in a sandwich ELISA. The mixture was allowed to incubate at room temperature for 30 minutes for the reaction to occur. The resulting solution was...

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Abstract

An antigen detection kit and an antigen detection method using the same are provided. The antigen detection kit comprises a capture antibody, a detection antibody bound to a single stranded DNA oligonucleotide, a single stranded RNA oligonucleotide complementary sequence to the DNA oligonucleotide, and an RNase.

Description

[0001] Cross-references to related applications: [0002] This application claims priority and benefit from Korean Patent Application No. 10-2009-0070041 filed with the Korean Intellectual Property Office on Jul. 30, 2009, the entire contents of which are incorporated herein by reference. Background of the invention: [0003] (a) Technical field [0004] The invention provides a kit for detecting an antigen, comprising a capture antibody, a detection antibody combined with a single-stranded DNA oligonucleotide, a single-stranded RNA oligonucleotide complementary to the DNA oligonucleotide, and RNase; And the method for detecting the antigen with the above kit. The invention adopts a single analysis means and a single RNase to realize the immunoassay of various biological materials. [0005] (b) Background technology [0006] ELISA (Enzyme-Linked Immunosorbent Assay) is an immunoassay that uses the reaction of a material with its corresponding antibody to detect the specific...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/53G01N33/68G01N33/52G01N21/64C12Q1/70C12Q1/68
CPCG01N2458/10G01N33/542G01N33/54306G01N33/533G01N33/563
Inventor 安大鲁杨恩卿韩基喆
Owner KOREA INST OF SCI & TECH
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