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MUC16的各个重组重复序列与CA125抗体的结合具有变异性。

Individual recombinant repeats of MUC16 display variable binding to CA125 antibodies.

作者信息

Wang Chien-Wei, Hanson Eliza K, Minkoff Lisa, Whelan Rebecca J

机构信息

Department of Chemistry, University of Kansas, Lawrence, KS, United States of America.

Department of Chemistry and Biochemistry, University of Notre Dame, Notre Dame, IN, United States of America.

出版信息

bioRxiv. 2023 Feb 9:2023.02.08.527749. doi: 10.1101/2023.02.08.527749.

DOI:10.1101/2023.02.08.527749
PMID:36798296
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC9934600/
Abstract

BACKGROUND

Despite its importance in the clinical management of ovarian cancer, the CA125 biomarker-located on the mucin protein MUC16-is still not completely understood. Questions remain about MUC16's function and structure, specifically the identity and location of the CA125 epitopes.

OBJECTIVE

The goal of this study was to characterize the interaction of individual recombinant repeats from the tandem repeat domain of MUC16 with antibodies used in the clinical CA125 II test.

METHODS

Using expression, we isolated nine repeats from the putative antigenic domain of CA125. Amino acid composition of recombinant repeats was confirmed by high-resolution mass spectrometry. We characterized the binding of four antibodies-OC125, M11, "OC125-like," and "M11-like"-to nine recombinant repeats using Western blotting, indirect enzyme-linked immunosorbent assay (ELISA), and localized surface plasmon resonance (SPR) spectroscopy.

RESULTS

Each recombinant repeat was recognized by a different combination of CA125 antibodies. OC125 and "OC125-like" antibodies did not bind the same set of recombinant repeats, nor did M11 and "M11-like" antibodies.

CONCLUSIONS

Characterization of the interactions between MUC16 recombinant repeats and CA125 antibodies will contribute to ongoing efforts to identify the CA125 epitopes and improve our understanding of this important biomarker.

摘要

背景

尽管CA125生物标志物（位于粘蛋白MUC16上）在卵巢癌临床管理中具有重要意义，但人们对其仍未完全了解。关于MUC16的功能和结构，尤其是CA125表位的身份和位置，仍存在疑问。

目的

本研究的目的是表征MUC16串联重复结构域中各个重组重复序列与临床CA125 II检测中使用的抗体之间的相互作用。

方法

通过表达，我们从CA125的假定抗原结构域中分离出九个重复序列。通过高分辨率质谱法确认重组重复序列的氨基酸组成。我们使用蛋白质印迹、间接酶联免疫吸附测定（ELISA）和局部表面等离子体共振（SPR）光谱法，表征了四种抗体（OC125、M11、“OC125样”和“M11样”）与九个重组重复序列的结合情况。

结果

每个重组重复序列都被不同组合的CA125抗体识别。OC125和“OC125样”抗体识别的重组重复序列不同，M11和“M11样”抗体识别的也不同。

结论

表征MUC16重组重复序列与CA125抗体之间的相互作用，将有助于正在进行的识别CA125表位的工作，并增进我们对这一重要生物标志物的理解。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/23be/9934600/2ee51df4e010/nihpp-2023.02.08.527749v1-f0004.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/23be/9934600/cabe6b679ad1/nihpp-2023.02.08.527749v1-f0001.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/23be/9934600/e1866d1e321d/nihpp-2023.02.08.527749v1-f0002.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/23be/9934600/cd356339f7e1/nihpp-2023.02.08.527749v1-f0003.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/23be/9934600/2ee51df4e010/nihpp-2023.02.08.527749v1-f0004.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/23be/9934600/cabe6b679ad1/nihpp-2023.02.08.527749v1-f0001.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/23be/9934600/e1866d1e321d/nihpp-2023.02.08.527749v1-f0002.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/23be/9934600/cd356339f7e1/nihpp-2023.02.08.527749v1-f0003.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/23be/9934600/2ee51df4e010/nihpp-2023.02.08.527749v1-f0004.jpg

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