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91.
92.
通过杂交瘤技术建立了3株抗鼠肺支原体单克隆抗体细胞株,它们分别是BA11,BD7和B612.试验表明:3株细胞所分泌的抗体均属IgG1亚类,都是鼠肺支原体的特异性抗体,与多种其它支原体无交叉反应。 相似文献
93.
本文比较了用HSV-1(IEA)和B病毒(IFA)的方法检查猴B病毒相关抗体和B病毒抗体的结果:IEA检查出的B病毒相关抗体阴性猴,经IFA检查,全部为B病毒抗体阴性。用IFA检查了B病毒相关抗体阴性的恒河猴,在单笼隔离饲养六个月后,有98.3%的动物B病毒相关抗体仍为阴性,表明IRA的阴性结果有较好的一致性。本文讨论了建立无B病毒感染猴群的可行性。 相似文献
94.
不同月龄婴儿接种DTP后血清中百日咳抗体水平的观测 总被引:2,自引:1,他引:1
本文对比观察了213例2月龄、3月龄婴儿接种DTP后百日咳抗体水平的变化,探讨了母传抗体对免后抗体增长的影响。结果表明2月龄、3月龄婴儿DTP免疫后1个月和3个月血清中百日咳抗体达保护水平的百分率无显著性差异(x ̄2=0.036,p>0.9;x ̄2=0.327,p>0.5),免后1个月抗体GMT无显著性差异(t=0.17,p>0.5),免后3个月抗体GMT3月龄组高于2月龄组(t=2.22,p<0.05)。我们还发现免前抗体水平与免后1个月GMT虽有负相关(r=—0,754)的倾向,但总体上对抑制免后抗体应答不明显,因而建议将儿童DTP基础免疫的起始月龄提前至2月龄进行。 相似文献
95.
We found an intra-acrosomal antigen of about 155,000 daltons (155 kDa) in a survey using the monoclonal antibody MC101 raised against mouse cauda epididymal spermatozoa. Morphological studies by means of indirect immunofluorescence and immunogold electron microscopy localized the antigen to the cortex region of the anterior acrosome. Avidin biotin complex immunocytochemistry initially demonstrated a faint signal at the anterior acrosome in the testis spermatozoa that increased in intensity as the sperm moved toward the distal epididymis. This incremental immunoreactivity was also confirmed by immunoblotting following one-dimensional SDS-PAGE. The 155 kDa protein band was immunostained, and it was much more intense in the cauda epididymal than in the caput and corpus epididymal spermatozoa. Only a trace or no immunostain was evident in the caput or testis spermatozoa. The antigen localization did not change during passage through the epididymis, being confined at the cortex region of the anterior acrosome. The epididymal epithelial cells were not immunostained. These findings suggested that the 155 kDa protein is biochemically modified, further implying that the biochemical alteration of intra-acrosomal material is involved in sperm maturation in the epididymis. © 1995 wiley-Liss, Inc. 相似文献
96.
Ralph Rapley 《Molecular biotechnology》1995,3(2):139-154
The exquisite specificity of monoclonal antibodies (MAb) has long provided the potential for creating new reagents for the
in vivo delivery of therapeutic drugs or toxins to defined cellular target sites or improved methods of diagnosis. However,
many difficulties associated with their production, affinity, specificity, and use in vivo have largely confined their application
to research or in vitro diagnostics. This situation is beginning to change with the recent developments in the applied molecular
techniques that allow the engineering of the genes that encode antibodies rather than the manipulation of the intact antibodies
themselves. Techniques, such as the polymerase chain reaction, have provided essential methods with which to generate and
modify the genetic constituents of antibodies, allow their conjugation to toxins or drugs, provide ways of humanizing murine
antibodies, and allow discrete modular antigen binding components to be produced. More recent developments of in vitro expression
systems and powerful phage surface display technologies will without doubt play a major role in future antibody engineering
and in the successful development of new diagnostic and therapeutic antibody-based reagents. 相似文献
97.
Sharfstein ST Tucker SN Mancuso A Blanch HW Clark DS 《Biotechnology and bioengineering》1994,43(11):1059-1074
Carbon-13 nuclear magnetic resonance (NMR) spectroscopy was used to study the metabolism of a murine hybridoma cell line at two feed glutamine concentrations, 4.0 and 1.7 mM. Carbon-13 labeling patterns were used in conjunction with nutrient uptake rates to calculate the metabolic fluxes through the glycolytic pathway, the pentose shunt, the malate shunt, lipid biosynthesis, and the tricarboxylic acid (TCA) cycle. Decreasing the feed glutamine concentration significantly decreased glutamine uptake but had little effect on glucose metabolism. A significant incrase in antibody productivity occurred upon decreasing the feed glutamine level. The increased antibody productivity in concert with decreased glutamine uptake and no apparent change in glucolytic metabolism suggests that antibody production was not energy limited. Metabolic flux calculations indicate that (1) approximately 92% of the glucose consumed proceeds directly through glycolysis with 8% channeled through the pentose shunt; (2) lipid biosynthesis appears to be greater than malate shunt activity; and (3) considerable exchange occurs between TCA cycle intermediates and amino acid metabolic pools, leading to substantial loss of (13)C label from the TCA cycle. These results illustrate that (13)NMR spectroscopy is a powerfulf tool in the calculation of metabolic fluxes, particularly for exchange pathways where no net flux occurs. (c) 1994 John Wiley & Sons, Inc. 相似文献
98.
Hamilton D Goodwin J Clarke MB du Moulin GC Liu V Caplan B Babbitt B 《Biotechnology and bioengineering》1994,43(8):700-705
An in vitro assay that measures the activation level of ex vivo activated (EVA) T cells currently being used in the adoptive immunotherapy of metastatic renal cell carcinoma has been developed. This assay is based on the ability of activated, but not resting. T cells to proliferate in response to the protein kinase C activator, phorbol myristate (PMA). To utilize this assay for in-process monitoring and control, we have begun an initial validation of the overall reproducibility of this assay. The proliferation of activated T cells in response to PMA, as measured by the mean cpm values of (3)H-thymidine incorporated, was demonstrated to have intra-assay coefficients of variation (cv's) for individual analysts that were typically less than 10% and rarely exceeded 20%. Activated T cells could be frozen and stored for at least 6 weeks with little or no deterioration in their ability to proliferate in response to PMA. Using these cells, inter-assay cv's that were typically less than 15% were obtained by individual analysts, and overall cv's of 10% to 25% were obtained for different samples assayed by different analysts at different times. This level of variability is very reasonable for a cellular assay. Furhter validation of this assay will address the issues of sensitivity, linearity and selectivity. To date, this assay has been used to analyze over 90 patient EVA cell samples and has revealed a broad range of proliferative responses to PMA. Taken together, these results suggest that this assay may be useful in defining the potency of the activated T cell used therapeutically. 相似文献
99.
Robinson DK Chan CP Yu Lp C Tsai PK Tung J Seamans TC Lenny AB Lee DK Irwin J Silberklang M 《Biotechnology and bioengineering》1994,44(6):727-735
Many mammalian cell fed-batch processes rely on maintaining the cells in a viable and productive state for extended periods of time in order to reach high final concentrations of secreted protein. In the work described herein, a nonamplified NSO cell line was transfected with a vector expressing a recombinant human anti-HIV gp 120 monoclonal antibody (Mab) and a selectable marker, glutamine synthetase. A fed-batch process was developed which improved product yields tenfold over the yields reached in batch culture. In this case, the clone was cultured for a period of 22 days and produced 0.85 g Mab/L. To gauge the effect of extended culture lifetime on product quality, biochemical characteristics of MAb isolated from different time points in the fed-batch culture were determined. The apparent molecular weight of the MAb was constant throughout the course of the culture. Isoelectric focusing revealed four major charged species, with a fifth more acidic species appearing later in the culture. The antigen binding kinetics were constant for MAb isolated throughout the culture period. Glycosylation analysis, on the other hand, revealed that MAb produced later in the culture contained greater percentages of truncated N-acetylglucosamine and highmannose N-glycans. Possible contributions to this underglycosylated material from either cell lysis or synthesis from noviable cells were found to be negligible. Instead, the viable cells appeared to be secreting more truncated and high mannose MAb glycoforms as the culture progressed. (c) 1994 John Wiley & Sons, Inc. 相似文献
100.
Effect of temperature on nucleotide pools and monoclonal antibody production in a mouse hybridoma 总被引:2,自引:0,他引:2
The specific monoclonal antibody productivity (q(Mab)) of a murine hybridoma (CC9C10) increased with incubation temperature in the range 33 degrees C to 39 degrees C. q(Mab) was constant at each temperature and was independent of the phase of culture. The q(Mab) increased 97% at 39 degrees C and decreased by 21% at 33 degrees C compared with controls at 37 degrees C. Specific rates of substrate (glucose and glutamine) utilization and byproduct (lactate and ammonia) formation also increased with temperature but the yield coefficient, Y(Lac/Llc') was constant for 33 degrees C to 39 degrees C and Y(Amm/Gin) was constant for 37 degrees C to 39 degrees C. Y(Amm/Gin) at 33 degrees C was lower than the control. Changes in specific nucleotide concentrations and ratios were monitored by analysis of intracellular nucleotide pools. The NTP ratio, (ATP + GTP)/(UTP + CTP), increased and the U-ratio (UTP/UDP-GNac) decreased during the course of each culture, whereas the adenylate energy charge, (ATP + 0.5ADP)/(ATP + ADP + AMP), remained relatively constant at a value 0.8. The relative content of UDP-/N acetyl galactosamine, UDP-N acetyl glucosamine, and NAD increased with incubation temperature, whereas the relative ATP content, SA(ATP + ADP + AMP)/SU (UTP + UDP-sugars) ratio, purine/pyrimidine, ATP/GTP, and U-ratio decreased at higher incubation temperatures. It is possible that these nucleotide parameters may have a regulatory role in the changes of q(Mab) observed at the higher temperatures. (c) 1994 John Wiley & Sons, Inc. 相似文献