聚乳酸和离聚物薄膜与重组抗体相结合用于处理性别分选牛精子的新见解。

New insights from poly-lactic acid and ionomer films coupled with recombinant antibodies for processing sexed-sorting bovine sperm.

作者信息

Thongkham Marninphan, Saenjaiban Aphisit, Jantanasakulwong Kittisak, Pattanawong Wiwat, Arjin Chaiwat, Hongsibsong Surat, Rachtanapun Pornchai, Sringarm Korawan

机构信息

Department of Animal and Aquatic Sciences, Faculty of Agriculture, Chiang Mai University, Chiang Mai 50200, Thailand.

Doctor of Philosophy Program in Nanoscience and Nanotechnology (International Program/Interdisciplinary), Faculty of Science, Chiang Mai University, Chiang Mai 50200, Thailand.

出版信息

Int J Biol Macromol. 2024 Jan;256(Pt 1):128425. doi: 10.1016/j.ijbiomac.2023.128425. Epub 2023 Nov 25.

DOI:10.1016/j.ijbiomac.2023.128425

PMID:38008136

Abstract

In this study, the efficacy of ionomers and poly-lactic acid (PLA) as an alternative solid material combined with scFv antibodies specific to bovine Y-sperm (Y-scFv) was studied to create a novel method of sexing technology. The coupling efficiency of Y-scFv to the surface of PLA, Na and Zn ionomer film was between 2 and 8 mg/mL. Fourier transform infrared spectra confirm that Y-scFv was bound with a carboxylic acid group in each film. Therefore, Na, Zn ionomers and PLA films conjugated with 4 and 8 mg/mL Y-scFv showed the highest concentration of Y-sperm in the eluted fraction. Considering that the elute fraction was enriched Y-sperm fraction, it contained 67.70-77.94 % of the Y-sperm ratio related to the produced supernatant fraction, which contained up to 69.31-76.01 % enriched X-sperm. In addition, the sperm quality after the sexing process was analyzed by CASA and imaging flow cytometry, which showed that each polymer did not have a negative effect on sperm motility and acrosome integrity for X-sperm. The capacity of ionomer and PLA combined with Y-scFv are used for bovine sperm sexing.

摘要

在本研究中，研究了离聚物和聚乳酸（PLA）作为替代固体材料与牛Y精子特异性单链抗体（Y-scFv）结合的效果，以创建一种新的性别鉴定技术方法。Y-scFv与PLA、Na和Zn离聚物膜表面的偶联效率在2至8 mg/mL之间。傅里叶变换红外光谱证实Y-scFv与各膜中的羧酸基团结合。因此，与4和8 mg/mL Y-scFv共轭的Na、Zn离聚物和PLA膜在洗脱组分中显示出最高浓度的Y精子。考虑到洗脱组分是富集的Y精子组分，其Y精子比例占产生的上清液组分的67.70 - 77.94%，而上清液组分中含有高达69.31 - 76.01%的富集X精子。此外，通过计算机辅助精子分析（CASA）和成像流式细胞术对性别鉴定过程后的精子质量进行了分析，结果表明每种聚合物对X精子的活力和顶体完整性均无负面影响。离聚物和PLA与Y-scFv结合的能力可用于牛精子性别鉴定。