西双版纳热带雨林凋落叶分解过程Ⅱ.微生物与线虫的群落动态

通过野外试验与室内模拟相结合,对西双版纳热带雨林生态系统混合凋落叶分解过程中微生物和线虫种群动态变化进行了系统研究。野外试验采用网袋法（即1mm和100μm网眼网袋,分别限制大型土壤动物和螨类的进入）,室内试验采用灭菌一接种法,以观测不同生物组成条件下线虫和微生物对凋落叶的分解作用。结果表明,伴随分解进程,微生物基本呼吸速率不断下降且与分解进程正相关。利用底物诱导呼吸法测定微生物生物量以及细菌、真菌生物量的变化表明,微生物在凋落叶分解过程中的演替遵循一定的路线。当土壤动物参与分解时,由于捕食压力的存在,微生物一般按照“双峰”型路线变化,并存在明显的生物演替现象,在这个过程中微生物对C的利用效率也不断提高;当不存在这种捕食压力时,微生物表现为“单峰-递减”式发展模式,生物量由强到弱变化,微生物对C的利用效率持续下降。土壤动物在凋落叶分解过程中表现为“单峰”型变化动态,与微生物量“双峰”动态形成互补,捕食者与被捕食者之间是一种“捕食-激发”作用下的种群消长关系,这种关系的强烈程度与捕食压力有关。“双峰”发展路线在一定程度上加速了凋落叶的分解进程。     

Marketing approval of mogamulizumab: A triumph for glyco-engineering

Therapeutic properties of antibodies strongly depend on the composition of their glycans. Most of the currently approved antibodies are produced in mammalian cell lines, which yield mixtures of different glycoforms that are close to those of humans, but not fully identical. Glyco-engineering is being developed as a method to control the composition of carbohydrates and to enhance the pharmacological properties of mAbs. The recent approval in Japan of mogamulizumab (POTELIGEO^®), the first glyco-engineered antibody to reach the market, is a landmark in the field of therapeutic antibodies. Mogamulizumab is a humanized mAb derived from Kyowa Hakko Kirin’s POTELLIGENT^® technology, which produces antibodies with enhanced antibody-dependent cell-mediated cytotoxicity (ADCC) activity. The approval was granted April 30, 2012 by the Japanese Ministry of Health, Labour and Welfare for patients with relapsed or refractory CCR4-positive adult T-cell leukemia-lymphoma.     

Interference of Thiomersal in Biologicals During Protein Estimation

In the present report thiomersal was detected as interfering substance in hepatitis B vaccines during the total protein estimation by Lowry's protein assay. The thiomersal at different concentrations of 0.005%, 0.0075%, 0.01%, 0.02%, 0.05% and 0.1% was found to reduce the Folin Ciocalteu's phenol reagent and produce colour development between 0.024 O.D to 1.023 O.D. values. Further, the thiomersal was shown to interfere between 34.55% to 52.73% with Folin Ciocalteu's phenol reagent, when 10 batches of different hepatitis B vaccines were subjected to estimation of total protein content by Lowry's protein assay.     

Testing the potency of anti‐TNF‐α and anti‐IL‐1β drugs using spheroid cultures of human osteoarthritic chondrocytes and donor‐matched chondrogenically differentiated mesenchymal stem cells

Inflammation plays a major role in progression of rheumatoid arthritis, a disease treated with antagonists of tumor necrosis factor‐alpha (TNF‐α) and interleukin 1β (IL‐1β). New in vitro testing systems are needed to evaluate efficacies of new anti‐inflammatory biological drugs, ideally in a patient‐specific manner. To address this need, we studied microspheroids containing 10,000 human osteoarthritic primary chondrocytes (OACs) or chondrogenically differentiated mesenchymal stem cells (MSCs), obtained from three donors. Hypothesizing that this system can recapitulate clinically observed effects of anti‐inflammatory drugs, spheroids were exposed to TNF‐α, IL‐1β, or to supernatant containing secretome from activated macrophages (MCM). The anti‐inflammatory efficacies of anti‐TNF‐α biologicals adalimumab, infliximab, and etanercept, and the anti‐IL‐1β agent anakinra were assessed in short‐term microspheroid and long‐term macrospheroid cultures (100,000 OACs). While gene and protein expressions were evaluated in microspheroids, diameters, amounts of DNA, glycosaminoglycans, and hydroxiproline were measured in macrospheroids. The tested drugs significantly decreased the inflammation induced by TNF‐α or IL‐1β. The differences in potency of anti‐TNF‐α biologicals at 24 h and 3 weeks after their addition to inflamed spheroids were comparable, showing high predictability of short‐term cultures. Moreover, the data obtained with microspheroids grown from OACs and chondrogenically differentiated MSCs were comparable, suggesting that MSCs could be used for this type of in vitro testing. We propose that in vitro gene expression measured after the first 24 h in cultures of chondrogenically differentiated MSCs can be used to determine the functionality of anti‐TNF‐α drugs in personalized and preclinical studies. © 2018 American Institute of Chemical Engineers Biotechnol. Prog., 34:1045–1058, 2018     

Affinity purification of a framework 1 engineered mouse/human chimeric IgA2 antibody from tobacco

Complex multimeric proteins such as dimeric and secretory immunoglobulin A (IgA) can be difficult to produce in heterologous systems, although this has been achieved using several platforms including plants. As well as topical mucosal applications, dimeric IgA (dIgA), and secretory IgA (sIgA) can be used in tumor and anti-viral therapy, where their more potent cell-killing properties may increase their efficacy compared to current drugs based on IgG. However, the development of therapeutic IgA formats is hampered by the need to co-express four different polypeptides, and the inability to purify such molecules using conventional protein A or protein G affinity chromatography. The light chain (LC)-specific affinity ligand protein L is a potential alternative, but it only recognizes certain kappa light chain (LC(κ)) subtypes. To overcome these limitations, we have adapted a framework-grafting approach to introduce LCs that bind protein L into any IgA. As a model, we used the chimeric anti-human chorionic gonadotropin (hCG) antibody cPIPP, since this contains a murine LC((κ)) subtype that does not bind protein L. Grafting was achieved by replacing selected framework region 1 (FR1) residues in the cPIPP LC(κ) variable domain with corresponding residues from LC(κ) subtypes that can bind protein L. The grafted antibody variants were successfully purified by protein L affinity chromatography. These modifications affected neither their antigen-binding properties nor the yields achieved by transient expression in tobacco plants. Our results therefore show that LC FR1 grafting can be used as generic strategy for the purification of IgA molecules.     

实验动物与兽用生物制品检验和研发

在兽用生物制品的质量检验和研发中,实验动物的应用极其普遍.其应用的历史悠久、使用面广,涉及的动物种类多、数量大.本文对应用动物进行兽用生物制品检验和研发中存在的问题进行了分析,并提出了建立大动物基地、保证供应,完善标准、保证质量,更新观念、完善法规、减少动物用量,加强管理、确保生物安全等方面的建议.     

美国生物制品中孤儿药的研发现状 及对中国孤儿药研发策略的思考

过去30 年, 美国在全球第一部《孤儿药法案》推动和相关政策法规引导下,生物科技飞速发展,尤其在孤儿药研发领域,成果卓著。 对这30 年来美国FDA 批准的生物制品中孤儿药产品作一解析,回顾美国对生物制品中孤儿药的研发策略,介绍孤儿药开发的新型商业 模式,同时反思中国孤儿药研发策略。     

A limulus amoebocyte lysate activating activity (LAL activity) that lacks biological activities of endotoxin found in biological products

Pyrogenic substances in influenza HA (IHA) vaccine have been controlled by the pyrogen test or the mouse body weight decreasing toxicity (BWD) test. We examined the possibility of replacing the animal tests with the endotoxin test Commercial IHA vaccines were found to show considerable levels of LAL activity ranging from 0.2 to 160 EU/ml. However, a batch of the vaccine having even 100 EU/ml of LAL activity showed neither pyrogenicity in rabbits nor tumor necrosis factor alpha (TNF-alpha) induction in RAW264.7 cells. The LAL activity of IHA vaccine was abolished by a monoclonal antibody that recognizes LPS-binding epitope of LAL factor C. The activity of IHA vaccine showed different physicochemical properties from those of LAL activity of endotoxin. LAL activity of endotoxin is known to be sensitive to polymyxin B treatment and was found to be resistant to polyoxyethylene 10 cetyl ether (Brij56) treatment. On the contrary, the LAL activity of IRA vaccine was shown to be resistant to polymyxin B but sensitive to Brij56 treatment. The difference in sensitivity of the two LAL activities to polymyxin B and Brij56 might suggest the possibility of their discriminative measurements.     

Biosimilars entering the clinic without animal studies: A paradigm shift in the European Union

The concept of biosimilars has spread from Europe to other regions throughout the world, and many regions have drafted regulatory guidelines for their development. Recently, a paradigm shift in regulatory thinking on the non-clinical development of biosimilars has emerged in Europe: In vivo testing should follow a step-wise approach rather than being performed by default. To not require animal testing at all in some instances can well be seen as a revolutionary, but science-based, step. Here, we describe the internal discussions that led to this paradigm shift. The mainstay for the establishment of biosimilarity is the pharmaceutical comparability based on extensive physicochemical and biological characterization. Pharmacodynamic comparability can be evaluated in in vitro assays, whereas pharmacokinetic comparability is best evaluated in clinical studies. It is considered highly unlikely that new safety issues would arise when comparability has been demonstrated based on physicochemical and in vitro comparative studies.