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Human lactoferrin monoclonal antibody pair

A monoclonal antibody and human lactoferrin technology, which is applied in the direction of anti-animal/human immunoglobulin, material inspection products, measuring devices, etc., can solve the problem of low sensitivity and specificity, which cannot meet the detection of human lactoferrin Requirements, poor matching, etc., to achieve high sensitivity and specificity

Active Publication Date: 2015-12-09
CHINESE ACAD OF INSPECTION & QUARANTINE
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] The use of double-antibody sandwich test paper for the detection of human lactoferrin can realize the rapid and easy detection of human lactoferrin; however, the existing anti-human lactoferrin antibodies have low sensitivity and specificity and poor pairing Poor, unable to meet the requirements of double-antibody sandwich immunoassay for detecting human lactoferrin

Method used

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  • Human lactoferrin monoclonal antibody pair
  • Human lactoferrin monoclonal antibody pair
  • Human lactoferrin monoclonal antibody pair

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0037] This example is used to illustrate the screening of human lactoferrin monoclonal antibody pairs.

[0038] The protein sequence of human lactoferrin (hLF) is shown in NCBIGenbank sequence number: U95626. According to the method in the literature (Penghua Yang, et al., Cattle Mammary Bioreactor Generated ya Novel Procedure of Transgenic Cloning for Large-Scale Production of Functional HumanLactoferrin., PLoSONE, 2008, 3(10), e3453), human lactoferrin (hLF) transgenic cows were obtained.

[0039] Intramuscular injection of prolactin needles (purchased from Xi'an Caotan Pharmaceutical Factory, the infertile cow lactation needles, the product is boxed, 10 No. I needles, 3 No. II needles) to transgenic dairy cows for lactation, of which No. I needles were injected intramuscularly continuously 10 days, once a day. One No. II needle was injected on the 13th, 15th, and 17th days respectively. From the 7th day of injection, the udders of the transgenic cows were massaged with w...

Embodiment 2

[0048] This example is used to illustrate the sensitivity and specificity of the monoclonal antibody of the present invention to the prepared immune colloidal gold test paper (ie, the immune colloidal gold test paper screened in Example 1) when testing actual samples.

[0049] (1) Sample preparation: collected milk, dissolved milk powder and liquid dairy products. Take about 200 μl of milk sample into the sample tube.

[0050] (2) Detection: Take out the immunocolloidal gold test strips screened in Example 1, equilibrate at room temperature for 20 minutes, open the package and take out the test strips, insert the test strips into the sample tube in the direction of the arrow, and let stand for 10-15 minutes ,judgement result.

[0051] (3) Judgment of results: At that time, there was a purple-red quality control line visible to the naked eye on the paper strip, but no visible purple-red detection line, it was judged as negative, and recorded as "-"; Control line, if there is ...

Embodiment 3

[0059] 140 milks known to be negative for human lactoferrin were collected (including 100 non-transgenic cows collected from a dairy farm in Dachang, Hebei, 20 transgenic cows with human lysozyme in a dairy farm at China Agricultural University and 20 human α-lactalbumin transgenic cows located in the dairy farm of China Agricultural University), and 22 known positive human lactoferrin milk (collected from the dairy farm located in China Agricultural University, transgenic human lactoferrin Gene cows), 162 samples in total, carried out blind test with the immune colloidal gold test paper obtained by screening in Example 1, and the measurement results showed that 140 known human lactoferrin-negative milks were all shown as negative for detection, and 22 known human lactoferrin Positive milk was all shown as positive for the test, and the diagnostic specificity of the test strip was 100%. Some of the results of the measurements can be seen in Figure 4 and Figure 5 shown.

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Abstract

The invention discloses a human lactoferrin monoclonal antibody pair, which is composed of a first monoclonal antibody and a second monoclonal antibody; wherein the first monoclonal antibody is secreted by a hybridoma cell strain with a preservation number of CGMCC No.10595, and the second monoclonal antibody is secreted by a hybridoma cell strain with a preservation number of CGMCC No.10594. The invention also provides an application method of the provided human lactoferrin monoclonal antibody pair in the preparation of a kit for detecting human lactoferrin. Through the technical scheme mentioned above, a double-antibody sandwich immunity test paper detection method is adopted to detect human lactoferrin, the sensitivity can reach 10 ng / mL; moreover, cross reaction does not happen between bovine lactoferricin and goat lactoferricin; cross section does not happen in human bacteriolysin transgene cattle and human alpha-serum albumin transgene cattle; and the sensitivity and specificity are high.

Description

technical field [0001] The present invention relates to the field of biotechnology, in particular to a human lactoferrin monoclonal antibody pair and its use. Background technique [0002] The advent of "Shuoshu" (Palmite et al., 1982) has accelerated the research on genetically modified organisms. The birth of the cloned sheep "Dolly" (Wilmute et al., 1997) has promoted the widespread application of transgenic technology in mammals. At present, a variety of transgenic pigs, cattle, and sheep (Schnieke et al., 1997; Toledo et al., 2006), which are used to produce medicinal or edible protein, increase lean meat percentage, improve nutrition, and enhance disease resistance, have been successfully developed. Their potential economic benefits It has huge social benefits and has good commercialization prospects. However, due to the risks of GMOs and their products, the management of GMOs and their products has always been highly valued by governments. [0003] In order to prot...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K16/18G01N33/68G01N33/577
Inventor 王勤刘晓飞林祥梅冯春燕李晓琳仇松寅刘丹丹
Owner CHINESE ACAD OF INSPECTION & QUARANTINE
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