Anti-CBir1 antibody detection test paper preparation method and purpose thereof

An antibody and detection strip technology is applied in the field of preparation of immunochromatographic detection test paper to achieve the effects of high detection performance, high sensitivity and high specificity

Inactive Publication Date: 2015-01-21
苏州和锐生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0009] In terms of IBD diagnosis, only the ELISA reagents of Promethues Laboratory in the United States and the chemiluminescent reagents developed by our company have been used commercially based on anti-CBir1 antibody detection reagents. Anti-CBir1 immunochromatographic test strips are still not available in the domestic and foreign markets. come out

Method used

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  • Anti-CBir1 antibody detection test paper preparation method and purpose thereof
  • Anti-CBir1 antibody detection test paper preparation method and purpose thereof
  • Anti-CBir1 antibody detection test paper preparation method and purpose thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0047] Example 1. Preparation of CBir1 antigenic protein

[0048] Antigen protein preparation method 1: Recombinant protein

[0049] 1) Reagent

[0050] Escherichia coli host bacteria DH5α, BL21(DE3), cloning vector pCR2.1T-vector, expression plasmid pET28a(+), DNA polymerase rTaq, T4 DNA ligase, DNA polymerase rTaq, LA Taq and restriction enzyme BamH I , Hind III and EcoR I, BamH I, DL2000 DNA Marker, T4 DNA ligase, low molecular weight standard protein, DNA gel recovery kit, IPTG, etc.

[0051] 2) Instrument

[0052] Ordinary shaker SCS-24; water-proof constant temperature electric heating incubator; Biophotometer spectrophotometer, desktop refrigerated centrifuge Centrifuge 5810R, desktop centrifuge MiniSpin; high-speed refrigerated centrifuge; protein electrophoresis and gel imaging system; PCR instrument; ultrasound Lysis instrument; constant temperature metal bath; HIS protein purification column, etc.

[0053] 3) Experimental method

[0054] a. Vector construction:...

Embodiment 2

[0062] Example 2 Antibody Preparation

[0063] Anti-human IgG monoclonal and polyclonal antibodies coated with antibodies for the conjugate pad and detection zone, as well as monoclonal and polyclonal antibodies from other animal sources for the quality control zone and conjugate pad, can be obtained by immunizing animals.

[0064] 1. The specific operation method of monoclonal antibody preparation is as follows:

[0065] The injection solution is obtained by mixing 0.05mg-5mg of immune preparations (respectively for goats, mice, rabbits, horses and guinea pigs) with 1 volume of Freund's complete adjuvant, and injects the injection solution into multiple parts of the animal skin;

[0066] One month later, the Freund's complete adjuvant mixture was subcutaneously injected into multiple parts of the animals to boost the immunization. After 7 to 14 days, the animals were bled, and the antiserum titers were measured.

[0067] Animals were boosted until titers plateaued. Hybrido...

Embodiment 3

[0073] Preparation of each component of the gold standard test paper of embodiment 3

[0074] 1. Preparation of colloidal gold solution

[0075] 0.01% (w / v) of HAuCl 4 Heat the solution to boiling, quickly add every 100mL HAuCl 4 Add an appropriate amount of reducing agent solution to the solution, and the color changes from blue, then light blue, to blue, then red after heating, and transparent orange-red after boiling for 7-10 minutes. Then filter with ultrafiltration or microporous membrane (0.45μM) to remove polymers and other impurities that may be mixed therein. The prepared colloidal gold should be pure, translucent, free of sediment and floating matter, and discard when oily matter and a large amount of black granular precipitate impurities appear on the liquid surface.

[0076] The reducing agent used therein can be trisodium citrate, tannic acid-trisodium citrate, white phosphorus, preferably trisodium citrate, more preferably 1% (w / v) trisodium citrate.

[0077]...

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Abstract

The invention discloses an anti-CBir1 antibody detection test paper, a preparation method and a purpose thereof. The test paper strip comprises a sample pad (1), a combination pad (2), an analysis membrane (3), a water absorption pad (6), a base plate (7), an inspection band line T (4) and a quality control band line C (5), wherein the analysis membrane is attached to the upper part of the base plate; the combination pad and the water absorption pad are respectively attached to two ends of the upper part of the analysis membrane; the sample pad is attached to one end of the upper part of the combination pad; and the inspection band line T and the quality control band line C are arranged on the analysis membrane. The CBir1 antigen protein is used for realizing rapid detection with high sensitivity and high specificity on the anti-CBir1 antibody in the sample, and provides basis for auxiliary diagnosis of autoimmune diseases such as clinic Crohn's disease.

Description

technical field [0001] The invention relates to the field of immunoassay detection. Specifically, the present invention relates to a preparation method and application of an immunochromatographic detection test paper for detecting CBir1. Background technique [0002] IBD is a common disease in North America and Europe. In the past 30 years, the incidence of IBD in Japan has gradually increased. Although there is no epidemiological data on the general population in my country, the number of patients with this disease has gradually increased in the past 10 years. . The clinical manifestations of IBD are complex and diverse, not only with gastrointestinal symptoms, but also with extraintestinal manifestations. Since the symptoms are non-specific enteritis manifestations such as abdominal pain, diarrhea, and blood in the stool, it is difficult to differentially diagnose CD and UC, as well as other intestinal chronic diseases such as IBD and tuberculous enteritis. [0003] Curr...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/68G01N33/531G01N33/532
CPCG01N33/6893G01N33/531G01N33/532G01N33/54306G01N2800/065
Inventor 陈菲霍如松周阳钱洋
Owner 苏州和锐生物科技有限公司
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