Optimized universal biological analysis method by adding monkey serum

A biological analysis and general-purpose technology, applied in the field of biological analysis, can solve the problems of high background value, affecting the sensitivity of the method, and unfavorable for the detection of animal exposure in low-dose groups, so as to optimize the method, improve the sensitivity and background value, and improve Effect of Optimizing Sensitivity

Pending Publication Date: 2020-05-22
WUXI APPTEC SUZHOU
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

This method can overcome the problem that the background value of general-purpose methods in monkey serum or monkey plasma will be relatively high, which will affect the sensitivity of the method and is not conducive to the detection of animal exposure in low-dose groups

Method used

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  • Optimized universal biological analysis method by adding monkey serum
  • Optimized universal biological analysis method by adding monkey serum
  • Optimized universal biological analysis method by adding monkey serum

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Experimental program
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Embodiment 1

[0031] 1. Experimental steps

[0032] It mainly includes the following steps:

[0033] 1) Prepare polyclonal antibody against human Fc segment and polyclonal antibody against human CH2 segment respectively, and use carbonate buffer to prepare 2 μg / mL coating buffer. Add 100 μL to each well and incubate overnight at 2-8°C.

[0034] 2) Add 300 microliters of PBS washing solution containing 0.5% Tween to each well, wash the plate 3 times and pat dry. Add 300 microliters of 3% BSA blocking buffer to each well and incubate at room temperature for 1-3 hours.

[0035] 3) Add 300 microliters of PBS washing solution containing 0.5% Tween to each well, wash the plate 3 times and pat dry. At the same time, drugs containing a certain gradient (5000, 2500, 1600, 800, 400, 200, 100, 50, 25, 10 ng / mL) were prepared in monkey serum, and 50-fold pre-diluted with 1% BSA. Then add 100 microliters of sample to each well, and incubate at room temperature for 2 hours at 400-600 rpm.

[0036] 4...

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Abstract

The invention discloses an optimized universal biological analysis method by adding monkey serum. The method comprises the following steps: preparing a universal coated antibody buffer solution, and carrying out incubating overnight; adding a closed buffer solution for incubation, adding two sets of same to-be-detected objects with certain gradients, and respectively adding the detection antibodyworking solution containing the monkey serum and the detection antibody working solution without the monkey serum into the to-be-detected objects; and comparing the background value and the sensitivity of each group of data after the data is processed by a microplate reader. According to the method, the problems that the sensitivity of the method is influenced and the detection of the exposure amount of animals in a low-dose group is not facilitated due to the fact that a general background value of a general method in monkey serum or monkey plasma is relatively high can be solved.

Description

technical field [0001] The invention relates to the field of biological analysis, in particular to reducing the background signal value to optimize the method and method sensitivity, and in particular to a method for optimizing general-purpose biological analysis by adding monkey serum. Background technique [0002] The investigation of sensitivity in macromolecular bioanalysis methods is an important indicator, and it is also a parameter that must be verified for bioanalysis methods currently recommended by FDA, NMPA, and OECD. The LLOQ as described in the FDA defines the sensitivity of the method and should be determined during method development. And develop and validate the sensitivity so that it can meet the requirements of the expected biological sample detection. The NMPA also describes sensitivity as the lowest concentration of analyte in a sample that can be reliably quantified with acceptable accuracy and precision. Sensitivity is the lowest point of the standard...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/68G01N33/543
CPCG01N33/6854G01N33/54393
Inventor 盛稳稳刘玮博金晓莺冀群升金毅施婧张欣
Owner WUXI APPTEC SUZHOU
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