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Process for preparing 2, 4-synthetic antigen with added spacing arms

A technology for synthesizing antigen and spacer arm, applied in the field of synthetic antigen preparation, can solve the problems of long time, complicated process, low specific antibody content, etc.

Inactive Publication Date: 2005-07-06
TSINGHUA UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The sample preparation (extraction, reduction, derivatization, and purification) process of the traditional detection method of 2,4-D is complicated and time-consuming, and the consistency of the detection results is poor, which limits the frequency and accuracy of routine detection
At present, the existing technology uses various cross-linking agents to directly condense the carboxyl group of the 2,4-D molecule and the amino group of the carrier protein molecule, and the molecules are connected by an amide bond, but part of the 2,4-D molecule on the synthetic antigen Masked by some long-chain groups on the surface of the protein, the content of 2,4-D specific antibodies produced by immunized animals is low, and the specificity and affinity are relatively weak

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0022] Example 1 is a method of adding a spacer arm by synthesizing a condensate of 2,4-D and a molecule having a C3 chain or a C6 chain (such as L-lysine), and then cross-linking with a carrier protein. The specific implementation steps are as follows:

[0023] 246.3mg of tert-butoxycarbonyl-lysine and 221mg of 2,4-D were dissolved in 10ml of PBS, and the pH was adjusted to 6.0, and 200mg of 1-ethyl-3-(3-dimethylaminopropyl)-dicarbonate was slowly added imine, reacted at 25°C for 2 hours.

[0024] Prepare the HCl / ethyl acetate solution by placing 200 g of solid NH in a 2 L suction filter flask 4 Cl, slowly add 98% concentrated H 2 SO 4 , the generated HCl gas is washed with sulfuric acid and CaCl 2 After drying in the drying tower, pour it into ice-cooled 500ml ethyl acetate, and seal the obtained solution in a refrigerator at 4°C.

[0025] The coupling product of 2,4-D and tert-butoxycarbonyl-lysine was dried with a rotary evaporator at 60°C, and the product was dissolv...

Embodiment 2

[0027] Example 2 is to use molecules (such as β-alanine, ε-aminocaproic acid or L-lysine) connected with C3 chain or C6 chain on the surface of the carrier protein, and then the carboxyl group of 2,4-D and act as A method for increasing the spacer by cross-linking the amino residues of the spacer molecule. The specific implementation steps are as follows:

[0028] Dissolve 121mg Tris and 150mg carrier protein (bovine serum albumin, rabbit serum albumin, chicken ovalbumin or hemocyanin) in 10ml PBS, adjust the pH to 6.0, slowly add 100mg-200mg 1-ethyl-3-( 3-Dimethylaminopropyl)-carbodiimide, and stir well. React at 25°C for 2 hours. Block the carboxyl group of the carrier protein.

[0029] Add 132 mg ε-aminocaproic acid (or 90 mg β-alanine) and 100 mg-200 mg 1-ethyl-3-(3-dimethylaminopropyl)-carbodiimide to the above reaction solution. Cross-link the carboxyl group of ε-aminocaproic acid or β-alanine with the amino group of the carrier protein. The reaction product solutio...

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PUM

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Abstract

The invention relates to a process for preparing a 2, 4 drop synthetic antigen with an increased partition arm, and belongs to micro-molecular environmental pollutant immune detection field. The process includes two methods: The first is synthesizing condensate of the 2, 4 drop and a molecule with a C3 chain or a C6 chain and cross linking it with a carrier protein. The second is linking molecules with C3 chains or C6 chains on surface of the carrier protein and condensing carboxy with amino acting as a partition arm molecule of the of 2, 4 drop. The invention can acquire the 2, 4 drop antibody with strong specificity and good affinity.

Description

technical field [0001] The invention belongs to the field of immunodetection of small molecular environmental pollutants, in particular to a preparation method of a synthetic antigen for the immunodetection of herbicide 2,4-D in water body and drinking water. Background technique [0002] Water contains a large number of harmful small molecular environmental pollutants, which is one of the main reasons why drinking water endangers human health. Extensive use of various synthetic pesticides (including 2,4-D) has led to the continuous increase of small molecule organic pollutants in surface water, making the water body lose its drinking function. The existing 2,4-D detection technology mainly utilizes liquid-solid extraction (Liquid-Solid Extraction) and gas chromatography-mass spectrometry (GC-MS). The sample preparation (extraction, reduction, derivatization, and purification) process of the traditional detection method of 2,4-D is complicated and time-consuming, and the co...

Claims

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Application Information

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IPC IPC(8): C07K1/107G01N33/531
Inventor 刘举施汉昌何苗余若祯
Owner TSINGHUA UNIV
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