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

A prostate-specific latex particle technology, which is applied in the field of clinical medical in vitro diagnostics, can solve problems affecting antigen binding and other problems, and achieve high correlation, wide application, and fast detection speed

Active Publication Date: 2014-08-27
BEIJING STRONG BIOTECH INC
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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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

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Experimental program
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Embodiment 1

[0049] Embodiment 1: Preparation of PSA monoclonal antibody

[0050] Human PSA was isolated from human semen plasma as described by Sensabaugh et al., 1990, J. Urology 144, 1523 . Mice were immunized with 4 injections of 25 μg human PSA in RAS (RIBI Adjuvant System) at regular time intervals. Four months after the last injection, lymphocytes isolated from the spleens of immunized mice were fused with the myeloma cell line SP2 / O-Ag14. Hybridoma cells secreting antibodies against PSA were identified by ELISA method. Multiple hybridomas with antibodies to different epitopes of PSA can be isolated. Then, the corresponding monoclonal antibodies were affinity purified, followed by epitope analysis. For epitope analysis, the surface plasmon resonance (SPR)-based biosensor BIAcore was used. Supernatants from cell cultures were tested, monoclonal antibodies were bound to the surface of the biosensor by polyclonal goat anti-mouse-Fc-antibodies, and the binding and dissociation of th...

Embodiment 2

[0051] Embodiment 2: preparation R1 reagent

[0052] Add sodium chloride 0.9%, PEG-60003%, BSA 0.5%, EDTA 50mM, and sodium azide 0.09% to 50mM HEPES buffer at pH 7.2, and stir evenly to form R1 reagent.

Embodiment 3

[0053] Embodiment 3: prepare PSA calibrator

[0054] In 50mM HEPES buffer solution with pH 7.2, add PSA with concentrations of 0, 4, 10, 23, 45, and 90ng / mL, and add sodium chloride 0.9%, BSA 1%, EDTA 50mM, sodium azide 0.1% , Stir evenly to be the 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 in vitro diagnosis of clinical medicine. Background technique [0002] Prostate specific antigen (PSA) is synthesized and secreted by prostate epithelial cells into seminal fluid, and is one of the main components of seminal plasma. In the late 1960s, Hare et al. found that a semen-specific protein with a molecular weight of about 34,000 was contained in prostatic fluid and semen during the study of immunocontraception. The protein was extracted and purified from prostate tissue in 1979. Because this protein is only present in prostate tissue, it was named prostate-specific antigen. [0003] There are two main forms of PSA in serum: one is free PSA (f-PS...

Claims

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Application Information

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Patent Type & Authority Patents(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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