Method for producing anti-human retinol conjugated protein antiserum for sheep

A protein-binding and retinol-binding technology, applied in the direction of anti-animal/human immunoglobulins, immunoglobulins from serum, etc., can solve the problems of low level, slow antibody production speed, high antigen dosage, etc. The effect of high level and small amount of antigen

Inactive Publication Date: 2015-10-21
海奥斯生物科技镇江有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although the current production methods can produce a certain amount of antibodies, the antibody production speed is slow, the level is low, and the amount of antigen is high

Method used

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  • Method for producing anti-human retinol conjugated protein antiserum for sheep

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0027] Choose 5 adult healthy goats, carry out the following steps:

[0028] (1) Antigen preparation: select antigen containing human retinol binding protein, and emulsify with Freund's adjuvant before injection to obtain antigen emulsion;

[0029] (2) The first injection: take 2.5mg human retinol binding protein antigen, dilute it to 5ml with PBS or normal saline, then add 5ml complete Freund's adjuvant to make antigen emulsion, use this antigen emulsion to inject sheep, The injection sites are multi-point subcutaneous injections on the soles of the feet, groins, armpits, necks, and backs. Each site has 1-2 points, totaling 10-16 points. equally distributed;

[0030] (3) Second injection: 20 days after the first injection, take the second injection, take 5.0 mg of human retinol binding protein antigen, dilute to 5 ml with PBS or normal saline, and then add 5 ml of incomplete Freund's adjuvant Make antigen emulsion, use this antigen emulsion to inject sheep, the injection si...

Embodiment 2

[0039] Select 5 adult healthy sheep and carry out the following steps:

[0040] (1) Antigen preparation: select antigen containing human retinol binding protein, and emulsify with Freund's adjuvant before injection to obtain antigen emulsion;

[0041] (2) The first injection: take 2.5mg human retinol binding protein antigen, dilute it to 5ml with PBS or normal saline, then add 5ml complete Freund's adjuvant to make antigen emulsion, use this antigen emulsion to inject sheep, The injection sites are subcutaneous multi-point injections on the soles of the feet, groins, armpits, necks, and backs. Each site has 1-2 points, a total of 10-16 points. The injection dose for each sheep is 2ml containing 0.5mg of antigen. equally distributed;

[0042] (3) Second injection: 20 days after the first injection, take the second injection, take 5.0 mg of human retinol binding protein antigen, dilute to 5 ml with PBS or normal saline, and then add 5 ml of incomplete Freund's adjuvant Make an...

Embodiment 3

[0051] Choose 5 adult healthy goats, carry out the following steps:

[0052] (1) Antigen preparation: select antigen containing human retinol binding protein, and emulsify with Freund's adjuvant before injection to obtain antigen emulsion;

[0053] (2) The first injection: take 2.5mg human retinol binding protein antigen, dilute it to 5ml with PBS or normal saline, then add 5ml complete Freund's adjuvant to make antigen emulsion, use this antigen emulsion to inject sheep, The injection sites are subcutaneous multi-point injections on the soles of the feet, groins, armpits, necks, and backs. Each site has 1-2 points, a total of 10-16 points. The injection dose for each sheep is 2ml containing 0.5mg of antigen. equally distributed;

[0054] (3) Second injection: 20 days after the first injection, take the second injection, take 5.0 mg of human retinol binding protein antigen, dilute to 5 ml with PBS or normal saline, and then add 5 ml of incomplete Freund's adjuvant Make an an...

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Abstract

The invention provides a method for producing anti-human retinol conjugated protein antiserum for sheep. The method comprises the following steps of: (1) preparing an antigen; (2) performing injection for the first time; (3) performing injection for the second time; (4) performing injection for the third time; (5) performing injection for the fourth time; (6) performing injection for the fifth time; (7) performing injection for the sixth time; (8) performing injection for the seventh time; (9) performing injection for the eighth time; and (10) detecting serum titer: drawing blood from the vein of the sheep after 8-10 days from the injection each time, separating the serum and detecting dual-extending titer. Compared with the prior art, the method for producing the anti-human retinol conjugated protein antiserum for the sheep, disclosed by the invention, is low in cost, and simple in process; the method not only can stably produce the antibodies, but also is high in antibody producing speed; the antibody level is high, and the dosage of the antigen is small.

Description

technical field [0001] The invention relates to a method for producing goat anti-human retinol-binding protein antiserum, which belongs to the technical field of antiserum preparation. Background technique [0002] Retinol-Binding Protein (RBP) is a vitamin transporter in the blood, synthesized by the liver, and widely distributed in blood, cerebrospinal fluid, urine and other body fluids. Determination of retinol-binding protein can detect renal tubular dysfunction early, and can sensitively reflect the degree of renal proximal tubule damage, and can also be used as an indicator of early liver function damage and monitoring and treatment. Its molecular weight is 21KD. [0003] At present, after emulsifying human RBP antigen and adjuvant, and immunizing animals according to certain procedures, anti-human RBP antibodies can be produced in animal serum. Although the current production methods can produce a certain amount of antibodies, the antibody production speed is slow, ...

Claims

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Application Information

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IPC IPC(8): C07K16/18C07K16/06
Inventor 张传明袁金城徐志伟
Owner 海奥斯生物科技镇江有限公司
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