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High-level expression of recombinant IgG1 by CHO K1 platform

Ningning Xu, Jianfa Ou, Al-Karim (Al) Gilani, Lufang Zhou, Margaret Liu

《化学科学与工程前沿（英文）》 2015年第9卷第3期页码 376-380 doi: 10.1007/s11705-015-1531-5

摘要： The Chinese Hamster Ovary (CHO K1) cell was used to express a targeted anti-cancer monoclonal antibody by optimizing the platform of the construction of production cell line in this study. The adherent CHO K1 was first adapted to suspension culture in chemical defined medium. Then the glutamine synthetase (GS) vector was applied to construct a single plasmid to overexpress a monoclonal antibody IgG1. Post transfection, the production of cell pool was optimized by glutamine-free selection and amplification using various concentrations of methionine sulfoximine. The best cell pool of CHO K1/IgG1 was used to screen the top single clone using the limiting dilution cloning. Finally, a high IgG1 production of 780 mg/L was obtained from a batch culture. This study demonstrated that the construction of high producing cell line, from gene to clone, could be completed within six month and the gene amplification improved protein production greatly.

关键词： Chinese hamster ovary (CHO) monoclonal antibody IgG1 amplification cell line development

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Control of lupus activity during pregnancy via the engagement of IgG sialylation: novel crosstalk betweenIgG sialylation and pDC functions

《医学前沿（英文）》 2023年第17卷第3期页码 549-561 doi: 10.1007/s11684-022-0965-7

摘要： Immunoglobulin (IgG) glycosylation affects the effector functions of IgG in a myriad of biological processes and has been closely associated with numerous autoimmune diseases, including systemic lupus erythematosus (SLE), thus underlining the pathogenic role of glycosylation aberration in autoimmunity. This study aims to explore the relationship between IgG sialylation patterns and lupus pregnancy. Relative to that in serum samples from the control cohort, IgG sialylation level was aberrantly downregulated in serum samples from the SLE cohort at four stages (from preconception to the third trimester of pregnancy) and was significantly associated with lupus activity and fetal loss during lupus pregnancy. The type I interferon signature of pregnant patients with SLE was negatively correlated with the level of IgG sialylation. The lack of sialylation dampened the ability of IgG to suppress the functions of plasmacytoid dendritic cells (pDCs). RNA-seq analysis further revealed that the expression of genes associated with the spleen tyrosine kinase (SYK) signaling pathway significantly differed between IgG- and deSia-IgG-treated pDCs. This finding was confirmed by the attenuation of the ability to phosphorylate SYK and BLNK in deSia-IgG. Finally, the coculture of pDCs isolated from pregnant patients with SLE with IgG/deSia-IgG demonstrated the sialylation-dependent anti-inflammatory function of IgG. Our findings suggested that IgG influences lupus activity through regulating pDCs function via the modulation of the SYK pathway in a sialic acid-dependent manner.

关键词： pregnancy IgG glycome type I interferon systemic lupus erythematosus

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Proteomics study of Mycoplasma pneumoniae pneumonia reveals the Fc fragment of the IgG-binding protein

《医学前沿（英文）》 2022年第16卷第3期页码 378-388 doi: 10.1007/s11684-021-0840-y

摘要： Macrolide and corticosteroid resistance has been reported in patients with Mycoplasma pneumoniae (MP) pneumonia (MPP). MP clearance is difficult to achieve through antibiotic treatment in sensitive patients with severe MPP (SMPP). SMPP in children might progress to airway remodeling and even bronchiolitis/bronchitis obliterans. Therefore, identifying serum biomarkers that indicate MPP progression and exploring new targeted drugs for SMPP treatment require urgency. In this study, serum samples were collected from patients with general MPP (GMPP) and SMPP to conduct proteomics profiling. The Fc fragment of the IgG-binding protein (FCGBP) was identified as the most promising indicator of SMPP. Biological enrichment analysis indicated uncontrolled inflammation in SMPP. ELISA results proved that the FCGBP level in patients with SMPP was substantially higher than that in patients with GMPP. Furthermore, the FCGBP levels showed a decreasing trend in patients with GMPP but the opposite trend in patients with SMPP during disease progression. Connectivity map analyses identified 25 possible targeted drugs for SMPP treatment. Among them, a mechanistic target of rapamycin kinase (mTOR) inhibitor, which is a macrolide compound and a cell proliferation inhibitor, was the most promising candidate for targeting SMPP. To our knowledge, this study was the first proteomics-based characterization of patients with SMPP and GMPP.

关键词： severe Mycoplasma pneumoniae pneumonia children proteomics Fc fragment of the IgG-binding protein mechanistic target of rapamycin kinase inhibitor

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IgG N-糖基心血管年龄独立于真实年龄精准表征心血管事件风险 Article

武志远, 郭政, 郑雨露, 王玉涛, 张海平, 潘慧颖, 李志伟, Lois Balmer, 李霞, 陶丽新, 郭秀花, 王嵬

《工程（英文）》 2023年第26卷第7期页码 99-107 doi: 10.1016/j.eng.2022.12.004

摘要：

亚临床动脉粥样硬化和代谢紊乱是心血管健康的重要风险因素，应用免疫球蛋白G（IgG）N-聚糖模式作为炎症指标表征其发病风险已有研究报道然而，对于IgG N-糖基谱在心血管疾病（CVD）风险分层中的能力仍然未知。本研究旨在利用IgG N-糖基标志物开发追踪心血管疾病风险的年龄指数。使用机器学习递归特征消除和惩罚回归算法逐步筛选特征糖基，并开发IgG N-糖基化心血管年龄（GlyCage）指数，以反映归因于心血管风险的与真实年龄间的偏差。因此，本研究开发的GlyCage指数利用IgG N-糖基谱追踪心血管健康水平。GlyCage和真实年龄之间的差距能够独立地表征心血管风险，提示IgG N-糖基化在心血管疾病的发病机制中起作用。

关键词： IgG N-糖基心血管年龄心血管年龄免疫球蛋白G 糖基化炎症特征选择机器学习

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Real time monitoring of bioreactor mAb IgG3 cell culture process dynamics via Fourier transform infrared

Huiquan Wu, Erik Read, Maury White, Brittany Chavez, Kurt Brorson, Cyrus Agarabi, Mansoor Khan

《化学科学与工程前沿（英文）》 2015年第9卷第3期页码 386-406 doi: 10.1007/s11705-015-1533-3

摘要： Compared to small molecule process analytical technology (PAT) applications, biotechnology product PAT applications have certain unique challenges and opportunities. Understanding process dynamics of bioreactor cell culture process is essential to establish an appropriate process control strategy for biotechnology product PAT applications. Inline spectroscopic techniques for real time monitoring of bioreactor cell culture process have the distinct potential to develop PAT approaches in manufacturing biotechnology drug products. However, the use of inline Fourier transform infrared (FTIR) spectroscopic techniques for bioreactor cell culture process monitoring has not been reported. In this work, real time inline FTIR Spectroscopy was applied to a lab scale bioreactor mAb IgG3 cell culture fluid biomolecular dynamic model. The technical feasibility of using FTIR Spectroscopy for real time tracking and monitoring four key cell culture metabolites (including glucose, glutamine, lactate, and ammonia) and protein yield at increasing levels of complexity (simple binary system, fully formulated media, actual bioreactor cell culture process) was evaluated via a stepwise approach. The FTIR fingerprints of the key metabolites were identified. The multivariate partial least squares (PLS) calibration models were established to correlate the process FTIR spectra with the concentrations of key metabolites and protein yield of in-process samples, either individually for each metabolite and protein or globally for all four metabolites simultaneously. Applying the 2 derivative pre-processing algorithm to the FTIR spectra helps to reduce the number of PLS latent variables needed significantly and thus simplify the interpretation of the PLS models. The validated PLS models show promise in predicting the concentration profiles of glucose, glutamine, lactate, and ammonia and protein yield over the course of the bioreactor cell culture process. Therefore, this work demonstrated the technical feasibility of real time monitoring of the bioreactor cell culture process via FTIR spectroscopy. Its implications for enabling cell culture PAT were discussed.

关键词： process analytical technology (PAT) Fourier-transform infrared (FTIR) spectroscopy partial least squares (PLS) regression mouse IgG3 bioreactor cell culture process real time process monitoring

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系统性红斑狼疮患者的血清IgG糖链特征 Article

潘胡丹, 王静蓉, 梁勇, 王灿坚, 田瑞敏, 叶华, 张晓, 吴沅皞, 邵苗, 张瑞军, 肖瑶, 李智, 张光峰, 周华, 王艺霖, 王晓双, 栗占国, 刘维, 刘良

《工程（英文）》 2023年第26卷第7期页码 89-98 doi: 10.1016/j.eng.2023.01.006

摘要：在本研究中，通过对389例SLE患者及304例健康对照者进行深入的糖组学分析，鉴定出血清免疫球蛋白G（IgG）上的两种N-糖链能够作为SLE的诊断生物标志物。基于片段特异性糖链分析和糖肽分析，发现这两种N-糖链生物标志物位于IgG上的Fc区域，并与疾病活动性密切相关。而酶学分析结果则提示，SLE患者体内一系列糖转移酶的失调可能是观察到的糖链产生变化的原因。研究结果为基于血清IgG糖基化的高效人群筛查和SLE潜在的新致病因素提供了新的思路。

关键词：系统性红斑狼疮 N-糖链诊断指标

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灰色预测模型GM（１，１）的适用性分析及在火灾风险预测中的应用

陈子锦,王福亮,陆守香

《中国工程科学》 2007年第9卷第5期页码 91-94

摘要：

通过对灰色预测模型———GM（１，１）的理论分析，证明了该模型的预测值及其变化趋势均具有单调性，进而提出了GM（１，１）模型的适用性判据，并给出了该判据在火灾风险灰色预测中的应用实例。

关键词：火灾预测 GM(1 1)模型火灾伤人率

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Metabolic interventions combined with CTLA-4 and PD-1/PD-L1 blockade for the treatment of tumors: mechanisms

《医学前沿（英文）》页码 805-822 doi: 10.1007/s11684-023-1025-7

摘要： Immunotherapies based on immune checkpoint blockade (ICB) have significantly improved patient outcomes and offered new approaches to cancer therapy over the past decade. To date, immune checkpoint inhibitors (ICIs) of CTLA-4 and PD-1/PD-L1 represent the main class of immunotherapy. Blockade of CTLA-4 and PD-1/PD-L1 has shown remarkable efficacy in several specific types of cancers, however, a large subset of refractory patients presents poor responsiveness to ICB therapy; and the underlying mechanism remains elusive. Recently, numerous studies have revealed that metabolic reprogramming of tumor cells restrains immune responses by remodeling the tumor microenvironment (TME) with various products of metabolism, and combination therapies involving metabolic inhibitors and ICIs provide new approaches to cancer therapy. Nevertheless, a systematic summary is lacking regarding the manner by which different targetable metabolic pathways regulate immune checkpoints to overcome ICI resistance. Here, we demonstrate the generalized mechanism of targeting cancer metabolism at three crucial immune checkpoints (CTLA-4, PD-1, and PD-L1) to influence ICB therapy and propose potential combined immunotherapeutic strategies co-targeting tumor metabolic pathways and immune checkpoints.

关键词： CTLA-4 PD-1 PD-L1 immune checkpoint blockade (ICB) metabolic reprogramming combined tumor therapeutic strategies

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渤海辽东带地质认识的突破与金县1-1大油田的发现

邓运华

《中国工程科学》 2011年第13卷第10期页码 13-18

摘要：在此认识指导下选择研究方向，评价有利目标，经钻探发现了金县1-1大油田，储量达1.5×10⁸ m³，充分显示了科研在油气勘探中的作用。

关键词：渤海辽东带金县1-1 地质认识大油田

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联邦无监督表示学习 Research Article

张凤达1,况琨1,陈隆1,游兆阳1,沈弢1,肖俊1,张寅1,吴超2,吴飞1,庄越挺1,李晓林3,4,5

《信息与电子工程前沿（英文）》 2023年第24卷第8期页码 1181-1193 doi: 10.1631/FITEE.2200268

摘要： FURL提出了两个新挑战：（1）客户端之间的数据分布转移（非独立同分布）会使本地模型专注于不同的类别，从而导致表示空间的不一致；（2）如果FURL中客户端之间没有统一的信息，客户端之间的表示就会错位。

关键词：联邦学习；无监督学习；表示学习；对比学习

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非等间距GM(1,1)幂模型及其工程应用

王正新,党耀国,刘思峰

《中国工程科学》 2012年第14卷第7期页码 98-102

摘要：

针对工程中大量存在的非等间距序列的建模问题，提出了非等间距GM(1, 1)幂模型。以平均相对误差绝对值最小化为目标，以模型参数之间的关系为约束，构建了一个非线性优化模型实现非等间距GM(1, 1)幂模型的参数估计。结果表明，非等间距GM(1, 1)幂模型的形式较为灵活，非等间距GM(1, 1)模型和灰色Verhulst模型均是非等间距GM(1, 1)幂模型的特殊情形，幂指数的优化有利于提高建模精度。最后通过一个工程实例验证了非等间距GM(1, 1)幂模型的有效性与实用性。

关键词：灰色系统非等间距序列 GM(1 1)幂模型参数优化

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磁控溅射法生长的带隙可调谐(GaxIn1−x)2O3 Research Articles

张法碧1,孙巾寓1,李海鸥1,周娟1,王荣1,孙堂友1,傅涛1,肖功利1,李琦1,刘兴鹏1,张秀云1,郭道友2,王相虎3,秦祖军1

《信息与电子工程前沿（英文）》 2021年第22卷第10期页码 1370-1378 doi: 10.1631/FITEE.2000330

摘要：采用磁控溅射技术和热退火技术在(0001)蓝宝石衬底上制备了多组分氧化物(GaxIn1−x)2O3薄膜，实现可调带隙。详细研究了三元化合物(GaxIn1−x)2O3在整个组成范围内的光学性质和能带结构演化。X射线衍射谱表明，Ga含量在0.11至0.55之间的(GaxIn1−x)2O3薄膜既有立方结构，也有单斜结构，而Ga含量高于0.74的(GaxIn1−x)2O3薄膜只有单斜结构。实验结果为透明导电化合物半导体(GaxIn1−x)2O3薄膜在光电和光伏行业的应用，特别是在显示器、发光二极管和太阳能电池的应用奠定了基础。

关键词： (GaxIn1−x)2O3薄膜；带隙可调谐；磁控溅射

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Soluble triggering receptor expressed on myeloid cell-1 (sTREM-1): a potential biomarker for the diagnosis

Changlin Cao, Jingxian Gu, Jingyao Zhang

《医学前沿（英文）》 2017年第11卷第2期页码 169-177 doi: 10.1007/s11684-017-0505-z

摘要： Sensitive and useful biomarkers for the diagnosis and prognosis of infectious diseases have been widely developed. An example of these biomarkers is triggering receptor expressed on myeloid cell-1 (TREM-1), which is a cell surface receptor expressed on monocytes/macrophages and neutrophils. TREM-1 amplifies inflammation by activating the TREM-1/DAP12 pathway. This pathway is triggered by the interaction of TREM-1 with ligands or stimulation by bacterial lipopolysaccharide. Consequently, pro-inflammatory cytokines and chemokines are secreted. Soluble TREM-1 (sTREM-1) is a special form of TREM-1 that can be directly tested in human body fluids and well-known biomarker for infectious diseases. sTREM-1 level can be potentially used for the early diagnosis and prognosis prediction of some infectious diseases, including infectious pleural effusion, lung infections, sepsis, bacterial meningitis, viral infections (e.g., Crimean Congo hemorrhagic fever and dengue fever), fungal infections (e.g., infection), and burn-related infections. sTREM-1 is a more sensitive and specific biomarker than traditional indices, such as C-reactive protein and procalcitonin levels, for these infectious diseases. Therefore, sTREM-1 is a feasible biomarker for the targeted therapy and rapid and early diagnosis of infectious diseases.

关键词： soluble triggering receptor expressed on myeloid cells-1 infectious diseases diagnosis and prognosis biomarker

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潘胡丹：风湿病IgG糖链结构改变及临床应用价值（2023年6月17日）

2023年06月29日

关键词：前沿技术与中医药创新发展

Molecular characterization of two suppressor of cytokine signaling 1 genes (

Xue XU,Jiannan ZHANG,Juan LI,Yajun WANG

《农业科学与工程前沿（英文）》 2015年第2卷第1期页码 73-83 doi: 10.15302/J-FASE-2015044

摘要： Suppressor of cytokine signaling 1 (SOCS1) protein can inhibit the signal transduction triggered by some cytokines or hormones and thus are important in many physiological/pathological processes, including innate and adaptive immunity, inflammation, and development in mammals. However, there is sparse information about their structure, tissue expression, in birds, where their biological functions remain unknown. In this study, we cloned and characterized two genes (named and ) from chickens. is predicted to encode a 207-amino acid protein, which shares high amino acid sequence identity (64%–67%) with human and mouse SOCS1. Besides , a novel gene was also identified in chickens and other non-mammalian vertebrates including . Chicken is predicted to encode a 212-amino acid protein, which shares only 30%–32% amino acid sequence identity with human SOCS1 and cSOCS1a. RT-PCR assay revealed that both and are widely expressed in all chicken tissues. Using a luciferase reporter assay system, we further demonstrated that transient expression of and can significantly inhibit chicken growth hormone (GH)- or prolactin (PRL)-induced luciferase activities of Hep G2 cells expressing cGH receptor (or cPRL receptor), indicating that SOCS1a and SOCS1b proteins can negatively regulate GH/PRL signaling. Taken together, these data suggest that both cSOCS1a and cSOCS1b may function as negative regulators of cytokine/hormone actions, such as modulation of GH/PRL actions in chickens.

关键词： chicken SOCS1a SOCS1b growth hormone prolactin