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Preprint ARTICLE | doi:10.20944/preprints202311.1302.v1

A ’Coomassie for Carbohydrates” – Novel Methodology for Rapid Detection and Quantitation of Glycosylated IgGs on an SDS-PAGE Gel

Arnab De, Christopher Sausen, Caryl Meade, Jing Zhou, Aaron D'Antona

Subject: Biology And Life Sciences, Life Sciences Keywords: glycosylation; monocloncal antobody; immunoglobulin

Online: 21 November 2023 (10:02:46 CET)

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Glycosylation is a common post-translation modification present on majority of eukaryotic proteins. Therefore, it is dependent on the host cell line and affected by the cell culture and purification process. Additionally, glycosylation of therapeutic biologics is known to have a profound impact on binding and downstream effector functions. Hence, glycosylation is increasingly considered a critical quality attribute (CQA) of biopharmaceutical drugs and has been showed to affect the safety and efficacy of these molecules. Therefore, regulatory agencies have adopted a Quality by Design (QbD) approach for glycan monitoring at various steps with stringent controls to ensure efficacy and safety. This requires multiple, orthogonal fit-for-purpose tools to study the process. One key unmet need in downstream organizations of large pharmaceutical companies is a real-time, rapid glycan detection and quantification tool to guide purification/expression process. To meet this need, we have developed a selective, colorimetric assay which selectively stains sugar moieties (glycosylated IgGs) which appears as magenta bands on an SDS-PAGE resolved gel. The selectivity arises because the method leverages stereochemical differences of hydroxyl groups between amino acids and carbohydrates to selectively stain the primary alcohol in carbohydrates but not in amino acids. Specifically, our method selectively oxidizes the ‘cis-diol’ group in carbohydrates to aldehydes and stains the aldehydes on an SDS-PAGE gel by Schiff’s reaction. Our method is semi-quantitative and has comparable sensitivity to Coomassie with LoD of around 50ng. The method can detect glycosylated IgGs produced in multiple cell lines (including HEK and CHO), thereby enabling us to study batch-to-batch variability and can critically stain both N-linked and O-linked glycosylation. We also show that the assay can be used as a downstream purification guide for process analytics following ion-exchange and hydrophobic interaction chromatography.

Preprint ARTICLE | doi:10.20944/preprints201711.0110.v1

Assessing the Effectiveness of a Far-red Fluorescent Reporter for Tracking Stem Cells In Vivo

Jing Zhou , Jack Sharkey, Rajeev Shukla, Antonius Plagge, Patricia Murray

Subject: Biology And Life Sciences, Biology And Biotechnology Keywords: Fluorescent reporter; E2-Crimson; mouse embryonic stem cells; knock-in; in vivo imaging

Online: 16 November 2017 (17:46:53 CET)

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Preprint ARTICLE | doi:10.20944/preprints202307.1991.v1

Digital Grading the Color Fastness to Rubbing of Fabrics Based on Spectral Reconstruction and BP Neural Network

Jinxing Liang, Jing Zhou, Xinrong Hu, Hang Luo, Genyang Cao, Liu Liu, Kaida Xiao

Subject: Computer Science And Mathematics, Artificial Intelligence And Machine Learning Keywords: textile fabrics; color fastness; digital grading; spectral reconstruction; BP neural network

Online: 28 July 2023 (10:42:44 CEST)

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Preprint ARTICLE | doi:10.20944/preprints201705.0008.v1

An Effective Surface Modification of Polyester Fabrics for Improving the Interfacial Deposition of Polypyrrole Layer

Zhenyun Zhao, Jing Zhou, Tao Fan, Lanqian Li, Zulan Liu, Yiping Liu, Ming Lu

Subject: Chemistry And Materials Science, Surfaces, Coatings And Films Keywords: polyester fabrics; surface modification; sulfuric acid; polypyrrole; deposition; conductivity

Online: 1 May 2017 (08:48:17 CEST)

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Preprint REVIEW | doi:10.20944/preprints201812.0156.v1

The Report of Marine Life Genomic Research

Guangyi Fan, Jianwei Chen, Tao Jin, Chengcheng Shi, Xiao Du, He Zhang, Yaolei Zhang, Hanbo Li, Ting Luo, Pengxu Yan, Guang Liu, Xiangqun Chi, Xiaoxuan Tan, Liangwei Li, Guilin Liu, Xiaochuan Liu, Shijie Hao, Kai Han, Xiaoyun Huang, Shuai Sun, Jing Zhou, Mengjun Yu, Lingfeng Meng, Yue Chang, Rui Zhang, Kaiqiang Liu, Mengqi Zhang, Yong Zhao, Chang Li, Jiao Guo, Xinyu Guo, Jiahao Wang, Meiqi Lv, Haoyang Gao, Yujie Liu, Yue Song, Shengjun Wang, Yang Deng, Binjie Ouyang, Jinzhong Lin, Yingjia Yu, lynn Fink, Xianwei Yang, Xun Xu, Xin Liu

Subject: Biology And Life Sciences, Ecology, Evolution, Behavior And Systematics Keywords: Marine microorganisms, marine fungi, algae, marine plants, marine invertebrates, marine vertebrates, genome, metagenome

Online: 12 December 2018 (15:44:09 CET)

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