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Blood group antibody detection method and application thereof

A blood group antibody and detection method technology, applied in application, chemical instruments and methods, botanical equipment and methods, etc., can solve the problems of inability to directly identify a single antibody and short storage period of identified spectrum cells.

Active Publication Date: 2022-03-01
BEIJING DAYOU TIANHONG TECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] At present, the detection technology of blood group antibodies mainly uses antibodies to identify spectrum cells. This method has problems such as short storage period of identification spectrum cells and inability to directly identify single antibodies. Therefore, the synthesis of recombinant proteins is very important for antibody identification.

Method used

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  • Blood group antibody detection method and application thereof
  • Blood group antibody detection method and application thereof
  • Blood group antibody detection method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0093] Example 1: Expression, purification and effect verification of Fya and Fyb blood group antibody binding proteins

[0094] 1. Expression of Fya, Fyb blood group antibody binding protein

[0095] In this example, the amino acid sequence (SEQ ID NO: 2) and the amino acid sequence (SEQ ID NO: 2) and The amino acid sequence of the Fyb blood group antibody binding protein (SEQ ID NO: 4), then construct the expression plasmid, amplify the target gene by PCR or outsource the synthetic gene sequence, and insert it into the pET32a vector through KpnI and EcoRI, after enzyme digestion and sequencing The size of the inserted gene fragment was identified and obtained. Among them, the expected inserted gene size of the Fya blood group antibody binding protein and Fyb blood group antibody binding protein expression plasmids was about 198bp, which was confirmed by enzyme digestion and was consistent with the expectation, and the sequencing result was correct.

[0096] After obtaining ...

Embodiment 2S

[0148] Example 2 Expression, purification and effect verification of S and s blood group antibody binding proteins

[0149] 1. Expression and purification of S and s blood group antibody binding proteins

[0150] In this example, the amino acid sequence (SEQ ID NO: 6) and amino acid sequence (SEQ ID NO: 6) and Amino acid sequence of s blood group antibody binding protein (SEQ ID NO: 8). Expression and purification of S and s blood group antibody binding proteins by eukaryotic expression vectors.

[0151] 1 Expression vector construction

[0152] Amplify the target gene by PCR or outsource the synthetic gene sequence; insert the target gene into the corresponding expression vector; sequence to obtain the plasmid containing the correct sequence; plasmid amplification production transfer downstream expression.

[0153] 2 Cell culture and protein expression

[0154] Use 293 serum-free CD medium (product number SMM293-TI) to subculture HEK293 cells, mix the target protein expre...

Embodiment 3

[0166] Example 3 Expression, purification and effect verification of N blood group antibody binding protein

[0167] 1. Expression and purification of N blood group antibody binding protein

[0168] In this example, the amino acid sequence (SEQ ID NO: 10) of the N blood group antibody binding protein was firstly designed according to the amino acid sequence of the N antigen coding region (SEQ ID NO: 9). The N blood group antibody binding protein was expressed and purified by eukaryotic expression vector.

[0169] The expression and purification steps of the N blood group antibody binding protein are consistent with the expression and purification steps of the S and s blood group antibody binding protein in Example 2. The final quality result is as Figure 17 As shown, the purified N blood group antibody binding protein and its molecular weight were obtained.

[0170] 2. Validation of the effect of N blood group antibody binding protein

[0171] Use the agglutination inhibi...

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Abstract

The invention provides a blood group antibody detection method, a blood group antibody detection kit, a blood group antibody binding protein, nucleic acid for coding the blood group antibody binding protein and a carrier containing the nucleic acid. Particularly, the blood group antibody binding protein comprises more than 80% of homology with SEQ ID NO: 2, 4, 6, 8 or 10 or comprises an amino acid sequence shown as SEQ ID NO: 2, 4, 6, 8 or 10, has antigen specificity, and can be used for blood group antibody detection.

Description

technical field [0001] The present invention relates to the technical field of blood group antibodies and antigens, in particular to a blood group antibody detection method and its application in the field of biomedicine. Background technique [0002] Duffy blood type is a blood group system that was discovered earlier. In 1950, Cusbush et al. found Fya antibody in the serum of a patient with multiple blood transfusions that caused hemolytic reactions. Since the patient’s surname was Duffy, they named this newly discovered blood type as Duffy blood type. A year later, Ikin et al. discovered the Fyb antibody corresponding to the Fya antibody. In 1995, the International Society of Blood Transfusion assigned the Duffy blood group system number 008. Fya antigen and Fyb antigen are fully developed when the fetus is born, and the existence of the antigen can be detected in the first 6-7 weeks of pregnancy. Duffy blood type has attracted much attention because of its important s...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/80G01N33/68G01N33/574G01N33/569G01N33/53C07K14/47C12N15/12
CPCG01N33/80G01N33/6893G01N33/574G01N33/56905G01N33/5308C07K14/47G01N2333/47G01N2800/24G01N2800/26G01N2800/7095
Inventor 朱思原
Owner BEIJING DAYOU TIANHONG TECH CO LTD
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