Acyl carrier protein III epitope, acyl carrier protein III antibody and use thereof

An acyl carrier protein, antigen epitope technology, applied in the fields of molecular biology and immunology, can solve the problem of not being able to trigger an immune response, and achieve the effects of easy spatial conformation reproduction, high specificity, and short fragments

Active Publication Date: 2013-08-28
BGI SHENZHEN CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0007] Although there are currently software for predicting antigenic epitopes, such as BEPITOPE software, about 30% of the predicted epitopes will be non-effective epitopes, that is, epitopes that cannot specifically represent antigens, or cannot Elicit immune response (Chang HT, LiuCH, Pai TW. Estimation and extraction of B-cell linear epitopes predicted by mathematical morphology approaches. J Mol Recognit. 2008 Nov-Dec; 21(6): 431-41)

Method used

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  • Acyl carrier protein III epitope, acyl carrier protein III antibody and use thereof
  • Acyl carrier protein III epitope, acyl carrier protein III antibody and use thereof
  • Acyl carrier protein III epitope, acyl carrier protein III antibody and use thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0044] Example 1: Prediction of Candidate Epitopes

[0045] The gene ID of the gene corresponding to rice acyl carrier protein Ⅲ in GenBank is: NP_001051948.1. The reading frame sequence is as follows:

[0046] ATGGCTGCTCCCATCACCGCCGCCACATCGCCTCTCTCCCCTGCGTCTCGGGTTCAGGTGATGTGCTCAATGCTCAATCCAACCTCAGCTTCTTTCTCCAGGCAAACAGCAAGCTTCCCGTCCATTCGGTTGCGCCCGGTCCCAAGTCGCTTCCAGGCCTTGTCTTGTTCGGCCAAACAAGACACTATTGATAAGGTTTGCGAAATAGTTAAGAATCAGTTAGCAGTAGATGAAGGCACTGCCGTTTCTGGAGAAACAAAATTCGTGGACCTTGGTGCTGATTCACTTGACACGGTTGAAATTGTGATGGGCCTTGAGGAGGCATTTCAGATTACCGTGGACGAATCGAGTGCGCAAGTGATTCAGACAGTGGAGGATGCTGCTGTGCTTATCGACAAGCTTGTGGCAGAGAAGGACGCCTAA(SEQ ID NO:1)

[0047] The full-length sequence of the encoded rice acyl carrier protein III is as follows:

[0048] MAAPITAATSPLSPASRVQVMCSMLNPTSASFSRQTASFPSIRLRPVPS TIDKVCEIVKNQLAVDEGTAVSGETKFVDLGADSLDTVEIVMGLEEAFQITVDESSAQVIQTVEDAAVLIDKLVAEKDA

[0049] (SEQ ID NO: 2, wherein the framed sequence is SEQ ID NO: 3)

[0050] Then, according to SEQ ID...

Embodiment 2

[0053] Example 2: Chemical Synthesis of Antigenic Epitopes

[0054] There is no cysteine ​​at both ends of the candidate antigen epitope obtained in Example 1. In order to realize the cross-linking with the carrier, the synthesized sequence needs to add cysteine, so the polypeptide sequence to be synthesized is: CRFQALSCSAKQD (SEQ ID NO :5).

[0055] The polypeptide sequence shown in SEQ ID NO: 5 was chemically synthesized (synthesized by Jill Biochemical Company) to obtain the antigenic epitope of acyl carrier protein III.

Embodiment 3

[0056] Embodiment 3: the preparation of epitope-KLH complex

[0057] Using the glutaraldehyde linkage method, the N-terminal of the epitope synthesized in Example 2 was cross-linked with the cross-linking carrier protein-keyhole limpet hemocyanin (KLH) to obtain the epitope-KLH complex.

[0058] The specific implementation steps are as follows:

[0059] Add 5 mg of synthetic peptide to 7 mg of KLH, slowly add 1 ml of freshly prepared 3 g / L glutaraldehyde solution while shaking, and incubate at room temperature for 2 h. The epitope-KLH complex was obtained by dialysis with pH 8.5 boric acid buffer for 24 hours.

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Abstract

The invention belongs to the field of molecular biology and immunology and relates to an acyl carrier protein III epitope, an acyl carrier protein III antibody and use thereof. Particularly, the acyl carrier protein III epitope has a sequence shown by SEQ ID No.3 or SEQ ID No.4 or SEQ ID No.5. The invention also relates to an acyl carrier protein III polyclonal antibody, which can be combined with the epitope specifically. The polyclonal antibody has high specificity and titer. The invention also relates to the preparation method and use of the polyclonal antibody, a composition containing the polyclonal antibody, and a method for detecting the acyl carrier protein III.

Description

technical field [0001] The present invention belongs to the fields of molecular biology and immunology. Specifically, it relates to an acyl carrier protein III epitope and an anti-acyl carrier protein III antibody. The present invention also relates to the composition respectively containing the antigen epitope or the antibody, and the use of the antibody in detecting acyl carrier protein III. Background technique [0002] Acyl carrier protein (ACP) is one of the components of the fatty acid synthase complex, involved in fatty acid synthesis. Phosphopantethein is the prosthetic group of ACP. Its phosphate group is connected with the serine residue of ACP by a phosphoester bond, and the -SH at the other end forms a thioester bond with the fatty acyl group. The molecule formed in this way is equivalent to a "swing arm" ", the substrate can be transferred from one catalytic center to another on the enzyme complex or from one enzyme reaction to another enzyme reaction. In pla...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07K7/08C07K14/795C07K14/765C07K14/47C07K16/18C07K16/06G01N33/577G01N33/53
Inventor 刘斯奇吴琳韦汉福曹英豪邱雪梅
Owner BGI SHENZHEN CO LTD
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