ELISA quantitative detection method of lactadherin or mucins 1 based on milk fat globule membrane characteristics

A quantitative detection method, milk fat globule membrane technology, applied in the field of biological detection, can solve the problems of difficult evaluation of solid phase carrier binding ability, difficulty of protein immune recognition, loss of MFGM protein, etc., achieving low cost, high accuracy rate, The effect of simplifying the detection steps

Active Publication Date: 2020-08-14
SHANGHAI CHILDRENS MEDICAL CENT AFFILIATED TO SHANGHAI JIAOTONG UNIV SCHOOL OF MEDICINE
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] (1) The MFGM protein may be partially or completely embedded in the membrane, which may cause difficulties for the immune recognition of the protein
[0007] (2) The process of sample processing may cause the loss of MFGM protein, and this error is unstable
[0008] (3) The binding ability of MFGM to solid phase support is difficult to evaluate, and proteins on the same MFGM may compete with each other
[0012] There is no report about the ELISA quantitative detection method of a milk agglutinin or mucin 1 based on the characteristics of the milk fat globule membrane of the present invention

Method used

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  • ELISA quantitative detection method of lactadherin or mucins 1 based on milk fat globule membrane characteristics
  • ELISA quantitative detection method of lactadherin or mucins 1 based on milk fat globule membrane characteristics
  • ELISA quantitative detection method of lactadherin or mucins 1 based on milk fat globule membrane characteristics

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0056] Example 1 Quantitative detection of lactoagglutinin

[0057] 1 Materials and methods

[0058] 1.1 Collection and packaging of breast milk

[0059] Randomly select 40 cases of breast milk from pregnant women undergoing full-term natural delivery or cesarean section in Shanghai Punan Hospital. All selected women are healthy women with no history of infectious diseases or chronic diseases such as hypertension, heart disease, etc. and no history of diseases during pregnancy . All selected women signed informed consent and voluntarily participated in the study. After obtaining maternal breast milk in Shanghai Punan Hospital, it was transferred to the laboratory in an ice box within 2 hours, and part of the breast milk was immediately selected and centrifuged (4°C, 3000g, 10 min) to obtain whey components. The whole milk and whey were divided into separate packages and stored at -80°C until use. During the process, the samples were homogenized and repeated freezing and tha...

Embodiment 2

[0094] Example 2 Quantitative detection of MUC1 protein

[0095] 1 main reagent

[0096] MUC1 ELISA kit (Thermo EHMUC1); native lysis buffer (ABCAM ab156035); TBST; 5*SDS sample buffer; antibody diluent; Running buffer; skim milk powder; primary antibody (Ab1): anti-MUC1 antibody (D908K, Cell Signaling).

[0097] 2 main consumables

[0098]Disposable Tip; absorbent paper; 1.5 / 2ml EP tube; shaker, incubation box, pipette, pipette gun, filter paper, nitrocellulose (NC) membrane.

[0099] 3 Experimental methods

[0100] 3.1 Collection and storage of breast milk samples

[0101] 3.1.1 Collection of breast milk samples

[0102] (1) Propaganda and education by professional medical personnel and signing the informed consent; 1 trained medical personnel for breast milk collection;

[0103] (2) Use a pump to collect breast milk once during the 42-day follow-up visit after delivery. First rinse the nipple with sterilized water, and collect all the milk from one breast.

[0104] ...

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Abstract

The invention relates to an ELISA quantitative detection method of lactadherin or mucins 1 (MUC1) based on milk fat globule membrane characteristics, and particularly relates to an enzyme-linked immunosorbent assay (ELISA) method. The method comprises the steps of collecting a breast milk sample, pretreating the breast milk sample by using a na ve lysis buffer, and placing the breast milk sample on ice for cracking, carrying out Western Blot quantitative detection, and carrying out ELISA quantitative detection. The invention further comprises a kit for quantitatively detecting the lactadherinor MUC1 in the breast milk sample. The kit comprises a na ve lysis buffer reagent and a specification. The method has the advantages that the method is used for quantitatively detecting the lactadherin or MUC1 in the breast milk of the puerpera in different stages, the flux is high, the accuracy is high, the detection steps are simplified, the cost is low, and a scientific basis is provided for improving the formula milk powder.

Description

technical field [0001] The invention relates to the technical field of biological detection, in particular to an ELISA quantitative detection method for lactagglutinin or mucin 1 based on the membrane properties of milk fat globules. Background technique [0002] Breast milk is the best food for babies, the only, safest and most complete natural food that babies need for growth and development. Breastfeeding can not only satisfy the baby's physical growth and development, but also promote intellectual development, which is beneficial to psychological growth; breastfed babies have good immune development, strong disease resistance, and "three highs" (hypertension, hyperlipidemia, High blood sugar) incidence is low; in addition, breastfeeding can reduce the incidence of breast cancer in mothers. Exclusive breastfeeding for newborns and infants is a scientific parenting method advocated by the World Health Organization and UNICEF, and it is the most basic measure to ensure the...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/68
CPCG01N33/68G01N2333/475G01N2333/4725
Inventor 陈同辛吴静李悦
Owner SHANGHAI CHILDRENS MEDICAL CENT AFFILIATED TO SHANGHAI JIAOTONG UNIV SCHOOL OF MEDICINE
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