ACA、AsAb与不孕及反复自然流产的关系

目的 探讨抗心磷脂抗体(ACA)、抗精子抗体(AsAb)与不孕、反复自然流产的关系.方法 采用酶联免疫吸附法(ELISA)对124例不孕患者(不孕组)、182例反复自然流产(流产组)及68例无流产史且已分娩的健康妇女(对照组)血清ACA及AsAb进行测定分析并比较.结果 不孕组ACA阳性率为45.16%,AsAb阳性率...     

取宫内节育器后继发不孕妇女血清免疫学检查结查分析

目的:探讨取宫内节育器(IUD)后继发不孕妇女血清中抗精子抗体(AsAb)、抗子宫内膜抗体(EMAb)、抗卵巢抗体(AoAb)和抗心磷脂抗体(ACA)的发生率及其作用,为临床病因分析和治疗提供依据。方法:采用酶联免疫吸附法(ELISA)对96例取IUD后继发不孕妇女、270例原发不孕妇女和50例正常生育未置IUD妇女(对照组)血清AsAb、EMAb、AoAb和ACA进行检测,并对检测结果进行统计分析。结果:取IUD后继发不孕妇女血清AsAb阳性32例(33.3%),EMAb阳性26例(27.1%),AoAb阳性21例(21.9%),ACA阳性25例(26.0%)其阳性率均显著高于对照组(P<0.01),同时AsAb和EMAb阳性率分别显著高于原发不孕组(P<0.01)。结论:取IUD后继发不孕妇女血清中AsAb、EMAb、AoAb和ACA阳性率显著升高,这可能是取IUD后继发不孕的一个重要原因。对取IUD后不孕妇女进行血清AsAb、EMAb、AoAb和ACA的检测具有一定意义。     

抗精子抗体对不孕不育和流产患者的影响分析

目的：探讨抗精子抗体（AsAb）对于不孕不育症以及流产患者的影响。方法收集不孕不育症以及反复自发性流产患者278例,并选择通同期健康体检者100例作为对照组,采用ELISA法测定两组的血清AsAb。结果男性不育患者的血清AsAb阳性率为22.35%,显著低于女性不孕患者的34.18%（P＜0.05）;不孕不育组的血清AsAb阳性率为30.04%,与流产组的28.57%无明显差异（P＞0.05）,但均显著高于对照组的2.00%（P＜0.05）;原发性不孕不育组的AsAb阳性率为31.05%,与继发性不孕不育患者的26.42%无明显差异（P＞0.05）;早期流产（孕周＜12周）者的AsAb阳性率为42.86%,显著高于晚期流产（≥12周）者的7.14%（P＜0.05）。结论AsAb可导致不孕不育症及反复自然流产和早期流产。     

抗HLA-I/II抗体、抗心磷脂抗体和抗β2糖蛋白1抗体与不孕和流产的关系

目的探讨抗HLA-I/II抗体、抗心磷脂抗体（ACA）和抗β2糖蛋白1（β2GP1）抗体与不孕和流产的关系。方法选取2011～2012年在本院就诊的不孕症患者550例、复发性流产患者1650例和正常分娩者150例。对各组分别进行抗HLA-I/II抗体、抗心磷脂抗体和抗β2GP1抗体检测,并将阳性率进行比较。结果不孕组抗HLA-I/II阴性率、ACA和抗β2GP1阳性率分别为76.0%、19.3%和17.5%;RSA组抗HLA-I/II阴性率、ACA和抗β2GP1阳性率分别为85.0%、23.5%和22.7%;均显著高于对照组,P〈0.05。结论抗HLA-I/II抗体、抗心磷脂抗体和抗β2GP1抗体是引起不孕和复发性流产的重要因素。     

反复自然流产患者血清P、AsAb、AcA和EmAb水平检测的临床意义

目的 探讨反复自然流产患者血清P、AsAb、AcA和EmAb水平的变化及意义.方法 应用放射免疫分析法和酶联法对32例反复自然流产患者进行血清P、AsAb、AcA和EmAb水平检测,并与35例正常妇女作比较.结果 反复自然流产患者血清孕酮(P)水平非常显著的低于正常人组(P<0.01).结论 血清P、AsAb、AcA和...     

复发性自然流产患者配偶精液分析及抗精子抗体的测定

目的 探讨复发性自然流产与患者配偶精子质量及抗精子抗原(AsAb)间的关系.方法 将256例诊断为复发性自然流产患者的配偶作为观察组,另选168例健康正常男性作为对照组.检查2组AsAb阳性率、精子密度、精子活力、畸形率测定.结果 观察组AsAb阳性率高于对照组,精子密度及精子活力低于对照组,差异均有统计学意义(P<0.01);精子畸形率较对照组高,特别是头部畸形率更高(P<0.01).结论 复发性自然流产患者配偶的精子活力、形态异常及抗精子复发性自然流产阳性率,可能与RSA的发生有一定关系.     

96例女性继发不孕患者免疫抗体检测结果分析

目的探讨抗精子抗体(AsAb)、抗子宫内膜抗体(EMAb)、抗心磷脂抗体(AcAb)检测在女性继发不孕患者中的临床应用价值。方法以96例女性继发不孕患者为观察组,同年龄阶段第一次怀孕健康正常女性186例为对照组,计算两组的AsAb、EMAb、AcAb阳性率,作统计分析。结果观察组AsAb、EMAb、AcAb阳性率分别是27.08%(26/96)、29.17%(28/96)、22.92%(22/96);对照组阳性率分别是3.23%(6/186)、1.08%(2/186)、2.15%(4/186);两组经统计比较P<0.01,有统计学意义。结论女性继发不孕患者的AsAb、EMAb、AcAb阳性率较正常健康第一次怀孕者明显增高,临床常规开展免疫抗体检测对女性继发不孕患者的诊断和治疗有积极意义。     

复发性自然流产患者配偶精液分析及抗精子抗体的测定

目的探讨复发性自然流产与患者配偶精子质量及抗精子抗原（AsAb）间的关系。方法将256例诊断为复发性自然流产患者的配偶作为观察组,另选168例健康正常男性作为对照组。检查2组AsAb阳性率、精子密度、精子活力、畸形率测定。结果观察组AsAb阳性率高于对照组,精子密度及精子活力低于对照组,差异均有统计学意义（P〈0.01）;精子畸形率较对照组高,特别是头部畸形率更高（P〈0.01）。结论复发性自然流产患者配偶的精子活力、形态异常及抗精子复发性自然流产阳性率,可能与RSA的发生有一定关系。     

276例AsAb、EmAb检测与不孕不育关系探讨

目的通过临床276例不孕患者抗精子抗体（AsAb）、抗子宫内膜抗体（EmAb）检测探讨其与不孕不育的关系。方法用ELISA法检测276例不孕不育患者AsAb、EmAb,和50例正常对照组对比研究。结果不孕妇女组AsAb阳性率26．1％（72／276）与正常对照组AsAb阳性率0．02％（1／50）比较χ2=14．13P〈0．01,差异有统计学意义。不孕妇女组EmAb阳性率为23．2％（64／276）与正常对照组EmAb阳性率0．0％（0／50）比较χ2=14．43P〈0．01,差异有统计学意义。结论AsAb和EmAb阳性能导致免疫性不孕。检测AsAb、EmAb对临床诊治不孕不育有重要价值。     

不孕不育患者血清中相关抗体检测临床研究

目的探讨自身免疫抗体血清学检测的意义及与不孕不育的相关性。方法将70例不孕不育患者作为观察组,另选健康体检者50例作为对照组。使用胶体金法检测2组患者血清中的抗精子抗体(AsAb)、抗子宫内膜抗体(EmAb)、抗心磷脂抗体(ACAb)、抗卵巢抗体(AoAb)。结果观察组AsAb、EmAb、ACAb、AoAb阳性率高于对照组;观察组男性AsAb阳性率为16.7%,低于女性的22.4%,差异均有显著统计学意义(P<0.01)。结论自身免疫抗体在不孕不育人群中有较高的阳性率。     

Antibody structure, instability, and formulation

The number of therapeutic monoclonal antibody in development has increased tremendously over the last several years and this trend continues. At present there are more than 23 approved antibodies on the US market and an estimated 200 or more are in development. Although antibodies share certain structural similarities, development of commercially viable antibody pharmaceuticals has not been straightforward because of their unique and somewhat unpredictable solution behavior. This article reviews the structure and function of antibodies and the mechanisms of physical and chemical instabilities. Various aspects of formulation development have been examined to identify the critical attributes for the stabilization of antibodies.     

评价自身抗体联合检测在诊断自身免疫性疾病中的应用

目的探讨检测抗核抗体（ANA）核型与抗可提取性核抗原（ENA）抗体及抗双链DNA（ds,DNA）抗体之间的相关性,提高对自身免疫性疾病（AID）的诊断及鉴别诊断参考价值。方法资料来源门诊及住院的AID病例血清标本进行检测,ANA检测采用问接免疫荧光法（IIF）,抗ENA抗体检测采用欧蒙斑点法（抗nRNP／SIn、Sm、SSA／Ro、SSB／Lo、Scl-70和Jo—1）六项抗体及抗（ds—DNA）抗体采用ELISA法检测。结果在98例ANA阳性病例血清标本中核型分布为：细胞核均质型（36．74％）,细胞核颗粒型（41．84％）,细胞核仁型（11．22％）,细胞核膜型（1．02％）,细胞核着丝点型（1．02％）;重叠核型中细胞核均质型／细胞核颗粒型（2．04％）,细胞核均质型／细胞浆颗粒型（6．12％）等。98例ANA与ENA同时阳性血清标本中42例抗双链DNA抗体阳性（42．86％）。36例均质型阳性样本中抗SSA／Ro抗体（24．48％）,抗SSB／Lo抗体（6．12％）,抗nRNP／Sm抗体（6．12％）。41例细胞核颗粒型阳性样本抗nRNP／Sm抗体（19．39％）,SSAfRo抗体（15．31％）,抗SSB／Lo抗体（6．12％）。ANA阳性可出现一种或多种自身抗体。结论ANA常见核型有：细胞核颗粒型,细胞核均质型及细胞核仁型。ANA均质型核型检出的抗ds-DNA抗体为主,且抗ds—DNA抗体含量与荧光强度成正比。ANA核型与抗ENA抗体二者之间有一定的相对应性。ANA及核型、抗ENA抗体和抗ds—DNA抗体的联合检测可明确诊断AID,而且利于判断患者血清中各种自身抗体与临床表现的关系。     

Antibody production

The clinical and commercial success of monoclonal antibodies has led to the need for very large-scale production in mammalian cell culture. This has resulted in rapid expansion of global manufacturing capacity [1], an increase in size of reactors (up to 20,000 L) and a greatly increased effort to improve process efficiency with concomitant manufacturing cost reduction. This has been particularly successful in the upstream part of the process where productivity of cell cultures has improved 100 fold in the last 15 years. This success has resulted from improvements in expression technology and from process optimisation, especially the development of fed-batch cultures. In addition to improving process/cost efficiencies, a second key area has been reducing the time taken to develop processes and produce the first material required for clinical testing and proof-of-principle. Cell line creation is often the slowest step in this stage of process development. This article will review the technologies currently used to make monoclonal antibodies with particular emphasis on mammalian cell culture. Likely future trends are also discussed.     

Antibody therapeutics, antibody engineering, and the merits of protein stability

Antibodies are highly soluble, multidomain proteins that are well suited for biopharmaceutical development; however, engineering antibodies to perform novel activities or to have enhanced clinical utility can have a detrimental effect on their biophysical properties. Various innovative designs, such as single-chain variable fragments (scFvs) and domain antibodies (dAbs), have been utilized to obtain the antigen-binding properties of natural antibodies, while using a minimal amount of the polypeptide sequence of an antibody. These designs can be used for generating diverse antibody libraries to support discovery and optimization and also serve as excellent building blocks for constructing more complex protein therapeutics, such as bispecific antibodies. However, engineered antibody-like proteins, including scFvs, are often unstable and prone to aggregation, compromising both protein production and quality. Research over the past few years has enhanced our understanding of how interdomain interactions within antibodies contribute to protein stability. This knowledge and sustained research to develop methods for modifying antibody fragments to improve stability have begun to have a positive impact on the quality of antibody libraries for discovery purposes and the viability of highly engineered proteins, such as bispecific antibodies, as therapeutics.     

Digoxin-Specific Antibody Fragments

Digitalis glycoside poisoning is an important clinical problem and the development of digoxin-specific antibody fragments (Fab) 30 years ago has changed clinical practice. Nevertheless, doubts still exist as to the appropriate dose indications for therapy. This paper reviews relevant literature, describes the difficulties associated with current treatment protocols and proposes an approach to therapy, which is based on theoretical principles and evidence gleaned from currently available clinical data sets. In patients with ‘acute’ poisoning, serum digoxin concentrations do not equate to the total body burden, as tissue distribution will not have occurred, and the calculations for present protocols, which use serum concentrations, are therefore likely to result in too much antibody being administered. Since a therapeutic quantity of digoxin will have little effect in a normal individual, complete neutralisation of all digoxin is also unnecessary. The pharmacokinetic and dynamic logic of using a smaller initial loading dose than predicted from total body calculations is rational. It is recommended that half the calculated loading dose, either based on serum concentration or history, should be administered and the impact on clinical features observed. If a clinical response is not seen within 1–2 hours, a further similar dose should be given. In the event of a full response, patients should be monitored for 6–12 hours; a second dose should only be given in the event of recurrence of toxicity. In patients with ‘chronic’ digoxin poisoning, the serum digoxin concentration will reflect the total body load. However, since such patients are invariably receiving digoxin for therapeutic purposes, full neutralisation is again not indicated. In addition, tissue redistribution of digoxin from deeper stores will occur following the binding of biologically active digoxin in the circulation. This process will occur over a number of hours and if the total calculated dose of antibody is administered in a single bolus, significant quantities will be excreted prior to redistribution of digoxin. Pharmacokinetic logic, therefore, suggests that half the calculated loading dose, based on serum concentration, should be administered and the impact on clinical features observed; a second dose should be given in the event of recurrence of toxicity.     

Monoclonal Antibody Therapy for Gram-Negative Sepsis: Principles,Applications, and Controversies

Gram-negative sepsis is a common event in hospitalized patients and is a leading cause of death in the United States. Endotoxin (lipopolysaccharide, LPS), a component of the cell wall of gram-negative microorganisms, is responsible for the cascade of events leading to the sepsis syndrome consisting of fever, tachycardia, tachypnea, and evidence of organ hypoperfusion. The lipid A region of endotoxin produces most of these biologic and toxic effects. Monoclonal IgM antibodies directed against the lipid A portion of endotoxin (anti-LPS MoAb) have been developed for the treatment of gram-negative sepsis. Results of two large-scale clinical trials suggest that these antibodies offer clinically and statistically significant reductions in mortality by a factor of about one-third. However, in both trials, this apparent beneficial effect was limited to particular subsets of patients, and no overall benefit was seen. These considerations, in addition to the likely high cost of the agents, pose questions about their ultimate use in the treatment of patients with gram-negative sepsis. Nevertheless, the logic of the approach, the demonstration of efficacy in disease models, and the advances in modern techniques of molecular biology all suggest that these or other closely related products will play a significant role in the treatment of this disorder.     

Population Pharmacokinetics of Sibrotuzumab, A Novel Therapeutic Monoclonal Antibody, in Cancer Patients

Population pharmacokinetics of sibrotuzumab, a humanized monoclonal antibody directed against fibroblast activation protein, were determined after multiple intravenous infusions of dosages ranging from 5 mg/m(2) to an absolute dose of 100 mg, in patients with advanced or metastatic carcinoma. In total, 1844 serum concentrations from 60 patients in three Phase I and II clinical studies were analyzed. The structural model incorporated two disposition compartments and two parallel elimination pathways from the central compartment, one linear and one nonlinear. Finally estimated pharmacokinetic parameters (%RSE) were: linear clearance CLL 22.1 ml/h (9.6), central distribution volume V1 4.13l (3.7), peripheral volume V2 3.19l (8.8), inter-compartmental clearance Q 37.6 ml/h (9.6); for the nonlinear clearance Vmax was 0.0338 mg/h (25) and Km 0.219 microg/ml (57). At serum concentrations between approximately 20 ng/ml and 7 microg/ml, the effect of the nonlinear clearance on pharmacokinetics was marked. Only at >7 microg/ml did CLL dominate overall clearance. Interindividual variability was 57% for CLL, 20% for V1 and V2, and 29% for Vmax and was larger than the inter-occasional variability of 13%. Of the many investigated patient covariates, only body weight was found to contribute to the population model. It significantly affected CLL, V1, V2 and Vmax resulting in marked differences in the model-predicted concentration-time profiles after multiple dosing in patients with low and high body weights. In conclusion, a robust population pharmacokinetic model was developed and evaluated for sibrotuzumab, which identified a possible need to consider body weight when designing dosage regimen for future clinical cancer trials.     

Antibody diversity singled out

Molecular biologist Michael Neuberger has been awarded the Royal Society GSK prize for his work on antibody diversification, highlighting the protective, as well as the destructive, role of DNA mutation.