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Preparation method of latex particles coated with prostate specific antigen-antibody and PSA enhanced turbidimetric immunophelometry kit

A prostate-specific, latex particle technology, used in the field of clinical medicine in vitro diagnosis, can solve problems such as affecting antigen binding, and achieve the effects of high correlation, wide popularization and application, and strong detection specificity

Active Publication Date: 2013-01-30
BEIJING STRONG BIOTECH INC
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Due to this adsorption force, some impurities are also adsorbed on the latex particles, which further affects the binding of the antigen (or antibody) to the particles.

Method used

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  • Preparation method of latex particles coated with prostate specific antigen-antibody and PSA enhanced turbidimetric immunophelometry kit
  • Preparation method of latex particles coated with prostate specific antigen-antibody and PSA enhanced turbidimetric immunophelometry kit
  • Preparation method of latex particles coated with prostate specific antigen-antibody and PSA enhanced turbidimetric immunophelometry kit

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0049] Example 1: Preparation of PSA monoclonal antibodies

[0050] Human PSA was isolated from human semen plasma as described by Sensabaugh et al., 1990, J. Urology 144, 1523. At certain time intervals, mice were immunized with 4 injections of 25 μg human PSA in RAS (RIBI Adjuvant System). Four months after the last injection, lymphocytes isolated from the spleen of the immunized mice were fused with the myeloma cell line SP2 / O-Ag14. Hybridoma cells secreting antibodies against PSA were identified by ELISA method. A variety of hybridomas with antibodies directed against different epitopes of PSA can be isolated. Then, the corresponding monoclonal antibodies were affinity purified and subjected to epitope analysis. For epitope analysis, the surface plasmon resonance (SPR)-based biosensor BIAcore was used. The supernatant of the cell culture is detected, the monoclonal antibody is bound to the surface of the biosensor by a polyclonal goat anti-mouse-Fc-antibody, and the bi...

Embodiment 2

[0051] Example 2: Preparation of R1 reagent

[0052] In 50mM HEPES buffer of pH7.2, add sodium chloride 0.9%, PEG-60003%, BSA 0.5%, EDTA 50mM, sodium azide 0.09%, and stir to obtain R1 reagent.

Embodiment 3

[0053] Example 3: Preparation of PSA calibrator

[0054] In 50mM pH7.2 HEPES buffer, PSA was added at concentrations of 0, 4, 10, 23, 45, 90ng / mL, and sodium chloride 0.9%, BSA 1%, EDTA 50mM, and sodium azide 0.1% were added. , stir evenly for PSA multi-point calibrator.

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Abstract

The invention relates to a preparation method of latex particles coated with a prostate specific antigen-antibody (PSA) and a PSA-enhanced turbidimetric immunophelometry kit. The latex particles coated with the PSA antibody are prepared by bonding the PSA antibody with latex particles by an optimized physical adsorption method. According to the invention, the kit utilizes the PSA antibody bonded onto the surface of the latex particles and the PSA in the human blood sample to conduct immunoreaction to form the turbidity, and the kit detects the content of the prostate specific antigen-antibody by the increase of the turbidity.

Description

technical field [0001] The invention provides a preparation method of latex particles coated with prostate specific antigen (PSA) antibody, and a kit for measuring PSA content in human blood samples by using an immunoturbidimetric method. The preparation method and the detection kit of the invention belong to the field of clinical medicine in vitro diagnosis. Background technique [0002] Prostate specific antigen (PSA) is synthesized and secreted into semen by prostate epithelial cells and is one of the main components of seminal plasma. At the end of the 1960s, Hare et al. found that there is a semen-specific protein with a molecular weight of about 34,000 in prostatic fluid and semen during the study of immunocontraception. This protein was extracted and purified from prostate tissue in 1979. Because this protein exists only in prostate tissue, it is named prostate-specific antigen. [0003] There are two main forms of PSA in serum: one is free PSA (f-PSA), which accou...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/531G01N33/577G01N21/82
Inventor 高爱民孙国敬张小锐徐丽刘希
Owner BEIJING STRONG BIOTECH INC
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