单克隆抗体可变区基因PCR扩增产物的直接测序

利用一组通用引物对单克隆抗体可变区ｃＤＮＡ进行ＰＣＲ扩增。扩增产物经纯化后通过一个以链终止法为基础的“循环测序”程序直接进行序列测定，其中所用四种双脱氧核苷酸分别带不同的荧光标记，测序引物选自同一组通用引物。测序电泳和数据收集在ＤＮＡ测序仪上进行。本法简单、快速、准确，是解析抗体可变区序列的有效手段。     

抗HSVgc单克隆抗体V_2基因的扩增、克隆及序列测定

本文从分泌具有高中和活性和高保护作用的抗ＨＳＶ－１／ＨＳＶ－２型共同性糖蛋白Ｃ的鼠源性单克隆抗体（ＭｃＡｂ）的杂交瘤细胞１Ａ１２中提取胞浆总ＲＮＡ，以Ｏｌｉｇｏ（ｄＴ）１５为引物逆转录合成ｃＤＮＡ，用一对鼠抗体Ｋ链通用引物进行ＰＣＲ，扩增出１Ａ１２ＶＬ基因，并克隆入ｐＵＣ１８质粒中。序列分析结果表明，１Ａ１２ＶＬ基因由３３０ｂｐ组成，编码１１０个氨基酸，隶属小鼠轻链ｋ链４组。     

T-Cell Subset Analysis by Monoclonal Antibodies in Primary Immunodeficiences

T-cell subsets have been analysed in 23 cases of primary immunodeficiency with monoclonal antibodies. Functional assays investigating T-cell function—proliferative response to mitogens, antigens, and allogeneic cells, cytotoxicity generated against allogeneic target cells and helper function to pokeweed mitogen-induced immunoglobulin synthesis—were performed in parallel in the same patients. The results enabled us to delineate four groups of patients. The first group consisted of patients in whom marker and function studies show concordant data, with either normal or strongly decreased T-cell number and function. The second group consisted of patients in whom various degrees of functional abnormality coexist with subnormal T-cell number and increased suppressor T-cell proportion. In the third group, we collected all patients who showed functional deficiencies without marker abnormalities. Finally, there was a small group composed of patients whose T-cell pattern was strongly suggestive of abnormal differentiation.     

地高辛单克隆抗体的研究 Ⅰ、分泌地高辛特异性单克隆抗体杂交瘤细胞系的建立及抗体特性分析

地高辛(Digoxin)是目前应用最广泛韵强心甙类药物,在心血管疾病临床治疗中占有十分重要的地位。强心甙治疗的安全范围较小,一般治疗量已接近中毒量的60%,并且有较大的个体差异,使得临床应用时中     

抗人CD3单链抗体基因的构建及序列分析

本文在已克隆抗人ＣＤ３抗体ＶＨ和ＶＫ基因的基础上，设计并合成了ＰＣＲ引物。两个外侧引物分别含有ＥｃｏＲＩ和ＳａｌＩ酶切位点及起始码和终止码序列，４个内侧引物各含部分连肽基因序列，回收后混合退火。     

利用表面等离子体共振技术测定BSA-抗BSA抗体间二维反应动力学

生物大分子间二维和三维反应动力学过程之间的本构关系还没有被完全认识.表面等离子体共振技术是成熟的研究三维反应动力学的方法.通过相同的技术实时记录耦联到载体(细胞、玻璃小球或脂质体)上的抗原分子与相应的耦联在芯片上的抗体分子之间,在不同实验条件下的反应动力学过程,建立了用表面等离子体共振技术测定抗体-抗原二维反应动力学的方法,可适用于建立生物大分子间二维与三维反应动力学间的定量关系.     

Analysis of Monoclonal Antibodies to Mycobacterium leprae by Crossed Immunoelectrophoresis

Monoclonal antibodies to Mycobacterium leprae were characterized in crossed immunoelectrophoresis and showed markedly different patterns of reactivity with M. leprae lines 2, 7 and 11 respectively. Line 7 corresponds to a cell wall-associated macromolecular complex containing lipid, polysaccharide, and two distinct 36 K and 65 K proteins.     

Redesignation of a Purported P1.15 Subtype-Specific Meningococcal Monoclonal Antibody as a P1.19-Specific Reagent

Two reference monoclonal antibodies against the meningococcal P1.15 subtype PorA, MN3C5C and 2-1-P1.15, showed only partial concordant recognition of meningococcal isolates. Cyanogen bromide cleavage of P1.19,15 PorA, peptide mapping, and sequencing of porA regions demonstrated that 2-1-P1.15 was specific for subtype P1.19, and henceforth it is to be redesignated as 2-1-P1.19.     

鼠抗HCV单克隆抗体研制及初步应用探讨

交联HCV 合成多肽抗原,用鼠一鼠杂交瘤技术获得8株稳定分泌抗HCV 的特异性单抗,分别命名为YLCV 系1—8。经初步研究结果表示,抗HCV 抗体阳性血清可抑制酶标YLCV-1和YLCY-5单抗与抗原的反应。抗原测定结果6份血清中有一份呈强阳性反应,由此提示单抗在HCV 抗体抗原检测系统中应用的可能性,同时为HCV 抗原成份的分析提供了必要的条件.     

两株新的抗肝细胞肝癌单克隆抗体的制备及特异性鉴定

以外科手术切除的肝癌标本制备的细胞悬液和新建株的肝癌细胞为免疫原。籍多重免疫途经致敢ＢＡＬＢ／ｃ小鼠，经２０次融合，从近１０，０００个融合孔中，筛选出两株（ＨＡｂ２５，ＨＡｂ２７）肝癌单克隆抗体，在９４例肝癌、１４１例其他肝病组织，２６种正常组织，１２９例各组织系统的肝外恶性肿瘤组织上做了交叉反应。除ＨＡｂ２７与毛细胆管有弱的反应外，两株抗体与肝癌组织结合的阳性率高（８３．１５％，８５．１１％），仅与少数消化道肿瘤有交叉．１３１Ⅰ标记抗体进行放射免疫显像，最佳显像时间为７２ｈ，瘤／肝、瘤／血比值分别为５．０７～６．８４、１．２６～２．１３，可望为肝癌导向药物研究，提供新的载体。     

金属螯合四肽与其单克隆抗体可变区相互作用的三维结构的计算机模拟

目的：模建两株抗金属螯合四肽单抗２Ｅ３－Ｆ７、１Ａ４－Ｄ１２的可变区与其半抗原分子的三维结构，并对它们之间相互作用的可能模式进行了分析、研究。方法：在ＳＧＩ图形工作站上，利用同源蛋白结构预测的方法模建分子结构，然后根据疏水性、静电性等将半抗原与抗体进行对接。结果：获得了两株抗体和半抗原的分子结构模型，并得到了它们可能的相互作用模式。结论：半抗原与抗体是能够合理结合的，而且这两株抗体具有酶催化活性的可能性。     

Characterization of Two Murine Monoclonal Antibodies Reactive with Human B Cells

We describe two monoclonal antibodies, HH1 and HH2. Both reacted selectively with surface immunoglobulin (sIg)-positive human B cells. Both antibodies stained on average 7-8% of peripheral blood mononuclear cells. They have not been found to react with cells or cell lines of other haematopoietic cell lineages, except that HH2 was positive on a small percentage of cells of the erythroid cell line K562. The molecular weight of the HH1 antigen was 95 kD, as established by Western blotting. Neither of these two antibodies reacted with Ig determinants, Fc receptors, complement receptors, or known class-I or class-II molecules. A combination of these antibodies was used in a direct panning technique for high-yield enrichment of normal B lymphocytes from peripheral blood. The enriched B cells could be further purified by lysis of T cells (final yield, on average 72 +/- 8% of initial B cells) or by a second panning (yield, 35 +/- 11%). The purified B cells contained less than 1% contaminating T cells and less than 0.5% monocytes and were used in an assay for B-cell-stimulating factor which they showed a normal and very reproducible proliferative response.     

青霉噻唑基单克隆抗体的研究——分泌青霉噻唑基单克隆抗体杂交瘤细胞株的建立

Kohler和Milstein于1975年创立的淋巴细胞杂交瘤技术已广泛地应用于生物学,医学等领域的研究,基于单克隆抗体的均一性,为用免疫化学方法检测药物及其中微量     

抗乙型肝炎病毒Pre S2 3B9单克隆抗体轻链可变区基因的扩增及序列分析

为了构建基因工程抗体并从分子水平分析抗乙型肝炎病毒ＰｒｅＳ２３Ｂ９单克隆抗体，我们利用分子生物学技术快速分离３Ｂ９杂交瘤细胞总ＲＮＡ，经反转录，ＰＣＲ扩增，分离获得了３Ｂ９单抗轻链可变区基因，并用聚合酶链反应（ＰＣＲ）技术及双脱氧核苷酸链末端终止法对该可变区基因的全部３２７ｂｐ核苷酸序列进行了测定。结果表明，３Ｂ９单抗轻链隶属小鼠免疫球蛋白Ｋａｐｐａ轻链第Ⅱ亚组，ＪＫ１微基因重排。     

Differentiation of Two Bovine Lentiviruses by a Monoclonal Antibody on the Basis of Epitope Specificity

Bovine immunodeficiency virus (BIV) and Jembrana disease virus (JDV) are bovine lentiviruses that are closely related genetically. A recombinant fusion protein containing the capsid protein of BIV expressed in Escherichia coli was used to immunize mice and produce monoclonal antibodies. Six hybridomas specific for BIV capsid protein were identified, and one antibody, designated 10H1, was characterized further. Competitive binding assays were performed to analyze the topography of antigenic determinants by enzyme-linked immunosorbent assay and demonstrated the existence of at least three distinct antigenic determinants on capsid protein. The monoclonal antibody reacted specifically with both BIV capsid and the recombinant fusion protein in Western immunoblot analyses. However, it did not react with the recombinant capsid fusion protein of JDV, indicating that BIV contains at least one unique epitope in the capsid protein that is absent in JDV. Further mapping of the epitope by chemical cleavage analysis identified that the epitope is located at the 6.4-kDa N terminus of the 29-kDa capsid protein. This monoclonal antibody assay will be valuable for distinguishing the two closely related lentiviruses by Western blotting.     

MG单抗相应抗原的理化特性分析

首先制备胃癌组织可溶性抗原提取液,经耐热试验,外源性凝集素结合试验,高碘酸氧化试验,甲醇—氯仿抽提试验等理化处理后,检测MG5,MG7,MG9,MGb1,MGe1,MGd1,6株单抗相应抗原的活性变化,进行初步定性分折。结果表明:MG单抗相应抗原均能被外源性凝集素ConA所识别,并可耐热(80℃30min,100℃20 mim),但高碘酸氧化后抗原活性则丢失。MG7,NGdl相应抗原可被甲醇-氯仿抽提,MG5,NG9,MGb1,MGe1相应抗原则不能被其抽提,故认为MG单抗相应抗原是一组碳水化合物抗原,其抗原决定族存在于糖链上,为抗原的进一步纯化和鉴定打下了基础。     

Cytochemical Analysis of Human Peripheral Blood Lymphocyte Subsets Defined by Monoclonal Antibodies

Human blood lymphocyte subpopulaiions, revealed by a panel of commercially available monoclonal antibodies by means of a resetting technique, were submitted to direct cyto-chemicat analysis and were shown to have distinguishing characteristics. T cells reactive with OKT3 antibody (T3⁺) displayed higher beta-glucuronidase and dot-like alpha-napnthyl acetate acid esterase (ANAE) activity than T3⁻ cells. Helper/inducer cells (T4⁺) were characterized by a high level of dot-like ANAE activity, whereas cytotoxic/suppressor cells (T8⁺) displayed selective naphthol-ASD-chloroacetate esterase activity. These results provide evidence for association of some cytoplasmic enzyme activities with the expression of membrane differentiation markers defined by monoclonal antibodies.     

抗人Ⅳ型胶原单抗研制及其特性

本文用胃蛋白酶提取的人胎盘Ⅳ型胶原免疫Balb/c小鼠,免疫脾脏细胞与小鼠骨髓瘤细胞融合后,经有限稀释法克隆获得两株分泌抗人Ⅳ型胶原单抗的杂交瘤细胞株CO_5和CO_6。用小鼠腹水和杂交瘤细胞培养上清纯化制备的两种单抗均属小鼠IgGl,K键,分别针对Ⅳ型胶原分子上的两个不同抗原决定簇。两种单抗特异性强,与人Ⅰ型、Ⅱ型、Ⅲ型、Ⅴ型、Ⅵ型、Ⅷ型胶原、Ⅳ型胶原7S区及人白蛋白等相关蛋白无明显交叉反应,且单抗效价高,亲和力强。