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Method for measuring glycine content in immunotoxin/antiserum

An assay method and antiserum technology, applied in the biological field, can solve problems such as no reports on related methods, and achieve the effects of simple derivation and good stability

Inactive Publication Date: 2012-07-18
玉溪九洲生物技术有限责任公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, the amino acid determination methods reported at home and abroad mainly include high-performance liquid chromatography (HPLC). Because HPLC has high sensitivity and specificity, the operation is relatively simple, and the requirements for equipment are moderate, but the determination of glycine in antiserum / antitoxin is related. The method has not been reported

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0021] Determination of glycine content in tetanus antitoxin:

[0022] Instruments: 1. Agilent 1200 high performance liquid chromatography and ChemStations workstation.

[0023] 2. Mettler AE 240 electronic balance.

[0024] Reagents and test drugs: reference substance glycine (National Institute for the Control of Pharmaceutical and Biological Products, batch number: 140689-200802, content: 100%); internal standard substance norvaline (Aladdin Chemistry, batch number: 17094, content: 99%); Phenylthiocyanate (DE, batch number: A0293365, content: 98%); triethylamine was chromatographically pure (TEDIA, batch number: 407145, content: 99.5%); n-hexane was analytically pure; water was ultrapure water; acetic acid Sodium is analytically pure.

[0025] Go through the following steps:

[0026] (1) Dissolve the internal standard norvaline in water to make an internal standard solution with a concentration of 3 mg / mL;

[0027] (2) Dissolve the reference substance glycine in water t...

Embodiment 2

[0033] Determination of glycine content in anti-rabies serum:

[0034] Instruments: 1. Agilent 1200 high performance liquid chromatography and ChemStations workstation.

[0035] 2. Mettler AE 240 electronic balance.

[0036] Reagents and test drugs: glycine reference substance (National Institute for the Control of Pharmaceutical and Biological Products, batch number: 140689-200802, content: 100%); internal standard norvaline (Aladdin Chemistry, batch number: 17094, content: 99%); Phenylthiocyanate (DE, batch number: A0293365, content: 98%); triethylamine was chromatographically pure (TEDIA, batch number: 407145, content: 99.5%); n-hexane was analytically pure; water was ultrapure water; acetic acid Sodium is analytically pure.

[0037] Go through the following steps:

[0038](1) Dissolve the internal standard norvaline in water to make an internal standard solution with a concentration of 4 mg / mL;

[0039] (2) Dissolve the reference substance glycine in water to make a re...

Embodiment 3

[0045] Determination of glycine content in antivenom serum:

[0046] Instruments: 1. Agilent 1200 high performance liquid chromatography and ChemStations workstation.

[0047] 2. Mettler AE 240 electronic balance.

[0048] Reagents and test drugs: Glycine reference substance (National Institute for the Control of Pharmaceutical and Biological Products, batch number: 140689-200802, content: 100%); internal standard α-aminobutyric acid (SIGMA, A2536, content: ≥99%); isosulfur Phenyl cyanate (DE, batch number: A0293365, content: 98%); triethylamine is chromatographically pure (TEDIA, batch number: 407145, content: 99.5%); n-hexane is analytically pure; water is ultrapure water; sodium acetate For analytical purity.

[0049] Go through the following steps:

[0050] (1) Dissolve the internal standard α-aminobutyric acid in water to make an internal standard solution with a concentration of 5 mg / mL;

[0051] (2) Dissolve the reference substance glycine in water to prepare a refe...

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PUM

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Abstract

The invention relates to a method for measuring glycine content in immunotoxin / antiserum. The method comprises the steps of: processing a sample in advance, removing macromolecular substances, utilizing isothiocyanato-benzene to conduct pre-column derivatization on the glycine, preparing reference solution after derivatization and sample supply solution after derivatization, and then conducting RP-HPLC (Reverse Phase High-Performance Liquid Chromatography). According to the method, the content of glycine in the immunotoxin / antiserum can be measured rapidly, simply and conveniently, so that the quality of a finished product can be evaluated.

Description

[0001] technical field [0002] The invention belongs to the field of biotechnology, and in particular relates to a method for determining glycine content in antitoxin / antiserum. Background technique [0003] Antitoxin / antiserum (Antitoxin / Antiserum) refers to the use of specific antigens to immunize animals, then collect high-titer immune serum, and after extraction and purification, prepare complete antibodies (IgG) or antibody fragments [F(ab') 2 ] immunoglobulin products. For the prevention and treatment of infections caused by the corresponding diseases. In 1890, Behring and Kitasato found that the serum of guinea pigs or rabbits injected with tetanus toxin could resist the infection and morbidity of tetanus after being injected into other animals. In 1891 in Berlin, Germany, the use of antiserum to successfully rescue a girl infected with diphtheria was the beginning of the use of antiserum in clinical treatment. Since then, many new varieties of animal immune sera ...

Claims

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Application Information

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IPC IPC(8): G01N30/89
Inventor 吴笛杨冬罗靖雄
Owner 玉溪九洲生物技术有限责任公司
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