Unlock instant, AI-driven research and patent intelligence for your innovation.

Detection method of protein content of antibody

A protein content and detection method technology, applied in the field of protein content detection, can solve the problems of long detection time, poor repeatability, multiple plate washing, etc., and achieve the effect of short detection time, low cost and few influencing factors

Inactive Publication Date: 2014-11-19
SICHUAN HUIYU PHARMA
View PDF1 Cites 0 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, this operation is more complicated, requires multiple washes, poor repeatability, and a long detection time, usually about 24 hours

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • Detection method of protein content of antibody
  • Detection method of protein content of antibody
  • Detection method of protein content of antibody

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0049] Rituximab (1.0 mg / ml) antibody samples were assayed.

[0050] Measured by protein chromatography system, using Mabselect SuRe 1ml prepacked column, gradient elution with mobile phase A and mobile phase B, the volume time of gradient elution is shown in Table 3, the injection volume is 500ul, and the flow rate is 1.0mL / min, the column temperature is 25°C, and the detection wavelength is 280nm;

[0051] The mobile phase A is a mixed aqueous solution of sodium dihydrogen phosphate dihydrate and 0.15M sodium chloride with a concentration of 20mM, and the pH is 7.2;

[0052] The mobile phase B is a mixed aqueous solution of 20 mM citric acid and 0.15 M sodium chloride, with a pH of 3.6.

[0053] The peak area of ​​the rituximab antibody sample was measured, and the concentration of the sample was calculated from the standard curve: y=164.71x+3.1171 to obtain the protein content of the antibody, as shown in Table 6.

[0054] Table 3 Volume schedule of mobile phase gradient...

Embodiment 2

[0057] Etanercept (1.0 mg / ml) antibody samples were assayed.

[0058] Measured with a protein chromatography system, using Mabselect SuRe 1ml prepacked column, gradient elution with mobile phase A and mobile phase B, the volume time of gradient elution is shown in Table 4, the injection volume is 400ul, and the flow rate is 0.5mL / min, the column temperature is 20°C, and the detection wavelength is 280nm;

[0059] The mobile phase A is a mixed aqueous solution of sodium dihydrogen phosphate dihydrate and 0.30M sodium chloride with a concentration of 10mM, and the pH is 7.0;

[0060] The mobile phase B is a mixed aqueous solution of 30 mM citric acid and 0.30 M sodium chloride, with a pH of 3.8.

[0061] The peak area of ​​the etanercept antibody sample was measured, and the concentration of the sample was calculated from the standard curve: y=164.71x+3.1171 to obtain the protein content of the antibody, as shown in Table 6.

[0062] Table 4 Volume schedule of mobile phase gr...

Embodiment 3

[0065] Cetuximab (1.0 mg / ml) antibody samples were assayed.

[0066] Measured by protein chromatography system, using Mabselect SuRe 1ml prepacked column, gradient elution with mobile phase A and mobile phase B, the volume time of gradient elution is shown in Table 5, the injection volume is 600ul, and the flow rate is 0.8mL / min, the column temperature is 30°C, and the detection wavelength is 280nm;

[0067] The mobile phase A is a mixed aqueous solution of sodium dihydrogen phosphate dihydrate and 0.05M sodium chloride with a concentration of 40mM, and the pH is 7.4;

[0068] The mobile phase B is a mixed aqueous solution of 50 mM citric acid and 0.05 M sodium chloride, with a pH of 3.4.

[0069] The peak area of ​​the cetuximab antibody sample was measured, and the concentration of the sample was calculated from the standard curve: y=164.71x+3.1171 to obtain the protein content of the antibody, as shown in Table 6.

[0070] Table 5 Volume schedule of mobile phase gradient...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

PUM

PropertyMeasurementUnit
concentrationaaaaaaaaaa
concentrationaaaaaaaaaa
Login to View More

Abstract

The invention discloses a detection method and in particular relates to a detection method of the protein content of antibody. The detection method comprises the following steps: (1) preparing a group of antibody standard substances with different concentrations, measuring the peak areas of the antibody standard substances by a protein chromatography system, and obtaining a standard curve according to the peak areas and the concentration gradient; (2) measuring the peak areas of antibody samples by the protein chromatography system, and calculating the concentrations of the samples according to the standard curve to obtain the protein content of the antibody. The method is easy to perform, good in repeatability, short in detection time and high in accuracy, has few influence factors, can be used for detecting the protein content of not only the purified antibody but also the antibody in cell culture liquid; moreover, different antibody standard substances can be selected and used to obtain the standard curve, and the protein contents of different antibody samples can be detected, so that the detection method is wide in application range.

Description

technical field [0001] The invention relates to a detection method, in particular to a detection method for the protein content of an antibody. Background technique [0002] Protein assay is a frequently used technique in life science research. At present, there are many methods for protein determination, but each has its limitations. The detection method of the protein content of conventional antibodies adopts UV280, that is, using a UV spectrophotometer to detect the OD value of the antibody at a wavelength of 280nm, and then calculate the protein content of the antibody according to the molar extinction coefficient of the antibody. This method can only detect the purified antibody, but cannot detect the protein content of the cell culture medium, because there are many additives, pigments, and impurities in the cell culture medium, which will interfere with the detection and lead to inaccurate detection results. [0003] In addition, the conventional Lowry method was us...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

Application Information

Patent Timeline
no application Login to View More
Patent Type & Authority Applications(China)
IPC IPC(8): G01N30/02
Inventor 刘玉应丁兆张仁怀胡勋雷富彬
Owner SICHUAN HUIYU PHARMA