Identification of B cell epitopes at the C-terminus of latency-associated nuclear protein of the kaposi's sarcoma-associated herpesvirus

The latency-associated nuclear antigen (LANA) of Kaposi's sarcoma-associated herpesvirus (KSHV) plays a key role in the induction of cell transformation, maintenance of viral episome, and modulation of immune response in human. To identify the presence of B cell epitopes within C-terminus of LANA and to characterize the monoclonal antibodies (MAbs) against this protein, we expressed the C-terminal region at aa 794-1000 of LANA (pLANA-C) in Escherichia coli as a fusion protein. KSHV-positive human sera were able to recognize the recombinant LANA-C in the Western blot analysis and ELISA. Mapping of antigenic epitopes of pLANA-C by KSHV-positive human sera revealed two B cell antigenic epitopes located at aa 846-854 and aa 794-822. The MAb 3F11 recognized a region between at aa 840 to 846 of LANA and exhibited a strong and specific binding to both pLANA-C and native viral LANA. These findings showed that pLANA-C and MAb 3F11 could be used for the detection of KSHV antibodies in human sera and for the advanced study of biological functions of LANA.     

应用于胶体金免疫层析的MDMA单克隆抗体的制备与鉴定

目的：制备高特异性的抗3,4亚甲二氧基甲基苯丙胺（MDMA）单克隆抗体,用于建立快速检测MDMA的胶体金免疫层析方法。方法：用MDMA人工抗原免疫BALB/c小鼠,将小鼠脾细胞与骨髓瘤细胞SP2/0融合,经亚克隆筛选得到稳定分泌抗MDMA单克隆抗体的杂交瘤细胞株。制备腹水,辛酸-饱和硫酸铵法纯化得到抗MDMA单克隆抗体（mAb）,并用胶体金免疫层析筛选出适用的抗MDMA单克隆抗体（mAb）,并对其进行特异性、纯度、亚类的鉴定分析。结果：经多次亚克隆后筛选得到4C10、4D10、7D11、8D4 4株分泌抗MDMA单克隆抗体的杂交瘤细胞株。其中细胞株8D4分泌的抗体适用于胶体金免疫层析。结论：成功筛选出能稳定分泌mAb的细胞株,为MDMA快速检测试剂的研制提供了关键材料。     

Analysis of the autoimmune epitopes on human testicular NASP using recombinant and synthetic peptides

The human nuclear autoantigenic sperm protein, NASP, is a testicular histone-binding protein of 787 amino acids to which most vasectomized men develop autoantibodies. In this study to define the boundaries of antigenic regions and epitope recognition pattern, recombinant deletion mutants spanning the entire protein coding sequence and a human NASP cDNA sublibrary were screened with vasectomy patients' sera. Employing panel sera from 21 vasectomy patients with anti-sperm antibodies, a heterogeneous pattern of autoantibody binding to the recombinant polypeptides was detected in ELISA and immunoblotting. The majority of sera (20/21) had antibodies to one or more of the NASP fusion proteins. Antigenic sites preferentially recognized by the individual patients' sera were located within aa 32-352 and aa 572-787. Using a patient's serum selected for its reactivity to the whole recombinant protein in Western blots, cDNA clones positive for the C-terminal domain of the molecule were identified. The number and location of linear epitopes in this region were determined by synthetic peptide mapping and inhibition studies. The epitope-containing segment was delimited to the sequence aa 619-692 and analysis of a series of 74 concurrent overlapping 9mer synthetic peptides encompassing this region revealed four linear epitopes: amino acid residues IREKIEDAK (aa 648-656), KESQRSGNV (aa 656-664), AELALKATL (aa 665-673) and GFTPGGGGS (aa 680-688). All individual patients' sera reacted with epitopes within the sequence IREellipsis.GGS (aa 648-688). The strongest reactivity was displayed by peptides corresponding to the sequence AELALKATL (aa 665-673). Thus, multiple continuous autoimmune epitopes in NASP involving sequences in the conserved C-terminal domain as well as in the less conserved testis-specific N-terminal region comprising the histone-binding sites, as predicted for an antigen-driven immune response, may be a target of autoantibodies in vasectomized men and may provide a relevant laboratory variable to describe more accurately the spectrum of autoantibody specificities associated with the clinical manifestation of vasectomy.     

Humoral immune response to functional regions of human cytomegalovirus glycoprotein B

Sera from patients with primary human cytomegalovirus (HCMV) infections, both acute and convalescent phase, and from HCMV-seropositive healthy subjects were analyzed to determine whether the sera would recognize antigenic domains on HCMV glycoprotein B (gB) that function in virion infectivity and spread of virus from cell to cell. The intact gB molecule, amino-terminal derivatives of different lengths, and internal deletion derivatives were expressed in eukaryotic cells and reacted by immunofluorescence with the sera. All convalescent-phase sera and most sera from healthy seropositive individuals reacted with full-length gB and with an amino-terminal derivative containing 687 amino acids (aa), gB-(1–687); approximately half of the sera recognized an amino-terminal derivative of 447 aa, gB-(1–447), and one-third reacted with the shortest deletion derivative of 258 aa, gB-(1–258). Of the acute-phase sera, 77% recognized intact gB and gB-(1–687), 32% recognized gB-(1–447), and 14% recognized gB-(1–258). Deletion of aa 548 to 618 dramatically reduced the percentage of reactive sera, whereas deletion of aa 411 to 447 had a minor impact on reactivity of sera. To investigate the epitope specificity of human antibodies to gB, we carried out competition experiments between human sera with neutralizing activity and selected monoclonal antibodies (mAbs) to conformational epitopes on gB. We found that antibodies in human sera preclude syncytium formation in UB15-11 glioblastoma cells constitutively expressing gB and compete with certain murine mAbs that block virus entry into cells and transmission of infection from cell to cell. Our results show that HCMV-immune human sera contain antibodies to functional regions on gB, and the abundance of these antibodies in convalescent-phase sera suggests that they may play a central role in limiting dissemination of virus in the host. J. Med. Virol. 52:451–459, 1997. © 1997 Wiley-Liss, Inc.     

Mapping of antigenic sites on the nucleocapsid protein of the severe acute respiratory syndrome coronavirus

Antigenic sites on the nucleocapsid (N) protein of severe acute respiratory syndrome (SARS) coronavirus (SARS-CoV) were mapped by Pepscan analysis with overlapping peptides that span the N protein sequence. Two major immunodominant epitopes located in the C-terminal region (amino acids [aa] 362 to 412) and middle region (aa 153 to 178) reacted with more than 75% of sera from SARS patients. Several minor immunodominant epitopes were reactive with about 50% of the SARS sera. Antisera from mice immunized with inactivated SARS-CoV recognized the two major immunodominant epitopes and one antigenic site located adjacent to the N-terminal region (aa 76 to 101), which did not react with the sera from SARS patients. Several monoclonal antibodies against SARS-CoV bound to the N- or C-terminal antigenic sites. These results suggest that the above antigenic sites on the N protein are important in eliciting humoral immune response against SARS-CoV in humans and animals and can be used as antigens for developing diagnostic tests.     

Analysis of antibody response to the measles virus using synthetic peptides of the fusion protein Evidence of non-random pairing of T and B cell epitopes

The measles virus induces a life-long immune response associated with antibodies specific for the fusion protein. To map the linear immunodominant recognition sites of the fusion (F) protein of the measles virus, we have reacted a complete set of 108 overlapping pentadecapeptides with purified IgG obtained from donor sera with elevated anti-measles titers. The antibodies recognized about 20% of the peptides and generated a characteristic binding pattern, defining about 6 or 7 distinctive regions (31–75; 111–145; 151–165; 191–215; 271–320; 421–440; 481–530) which include the major hydrophobic segment (111–145) of the intersubunit region and the C-terminal Cys-cluster region. The binding sites were located in close proximity of the few experimentally defined T cell epitopes. This pairing of T and B cell epitopes was corroborated by computer-assisted T cell prediction. The significance of a non-random association of T and B cell epitopes for processing and presentation is discussed. It is speculated that in long-term immunity against measles (F protein), B cells of the same sIg specificity play an important role both as antigen presenting cells and as antibody producing cells. In contrast to human sera from late convalescent donors, mouse and rabbit MV antisera with high neutralizing titers as well as neutralizing MV-F specific monoclonal antibodies did not react with the peptides.     

A peptide mimotope of hepatitis C virus E2 protein is immunogenic in mice and block human anti‐HCV sera

Conformational B‐cell epitopes on the HCV E2 protein recognized by human antibodies were characterized by the use of a peptide mimotope named K1. K1 was identified by two HCV anti‐E2 monoclonal antibodies (mAbs) following selection and purification of phage clones containing a 15‐mer random peptide insert. Murine antisera to the mimotope K1 recognized the E2 protein. Five of eight human sera from patients who had cleared HCV recognized the K1 mimotope. Binding to E2 in four individuals with the capacity to block E2–CD81 interaction was inhibited by the mimotope K1. The results demonstrate that anti‐E2 antibodies in sera from patients who have cleared HCV infection are directed against a conformational B‐cell epitope on E2 that can be mimicked with linear synthetic peptides. These findings could have implications for vaccine design by employing linear mimotopes to direct B‐cell responses against those specific E2 epitopes that may correlate with immunity. J. Med. Virol. 82:1655–1665, 2010. 2010 Wiley‐Liss, Inc.     

Characterization of Zaire ebolavirus glycoprotein-specific monoclonal antibodies

Zaire ebolavirus (ZEBOV) can be transmitted by human-to-human contact and causes acute haemorrhagic fever with case fatality rates up to 90%. There are no effective therapeutic or prophylactic treatments available. The sole transmembrane glycoprotein (GP) is the key target for developing neutralizing antibodies. In this study, recombinant VSVΔG/ZEBOVGP was used to generate monoclonal antibodies (MAbs) against the ZEBOV GP. A total of 8 MAbs were produced using traditional hybridoma cell fusion technology, and then characterized by ELISA using ZEBOV VLPs, Western blotting, an immunofluorescence assay, and immunoprecipitation. All 8 MAbs worked in IFA and IP, suggesting that they are all conformational MAbs, however six of them recognized linearized epitopes by WB. ELISA results demonstrated that one MAb bound to a secreted GP (sGP 1-295aa); three bind to a part of the mucin domain (333-458aa); three MAbs recognized epitopes on the C-terminal domain of GP1 (296-501aa); and one bound to full length GP (VLPs/GP1,2 ΔTm). Using a mouse model these MAbs were evaluated for their therapeutic capacity during a lethal infection. All 8 MAb improved survival rates by 33%-100% against a high dose lethal challenge with mouse-adapted ZEBOV. This work has important implications for further development of vaccines and immunotherapies for ZEBOV infection.     

Mapping of the region predominantly recognized by antibodies to the Plasmodium falciparum merozoite surface antigen MSA 1.

The locations of the epitopes of a panel of mouse monoclonal antibodies directed against the Plasmodium falciparum merozoite surface antigen MSA 1 were mapped by using naturally occurring processed fragments, by chemical cleavage of the protein and by comparison of the isolate-specificity of binding with known sequence variation. By these criteria, the most antigenic region occurs in the cysteine-rich, invariant 19-kDa carboxyl terminal domain with 12/19 monoclonal antibodies (mAbs) binding to this region. One of these mAbs recognized an epitope near the C-terminal putative glycosylphosphatidylinositol anchor site. This was the only mAb which significantly inhibited parasite growth in vitro. The other mAbs recognized conformational epitopes involving the cysteine residues located throughout this fragment. This study has identified further naturally occurring processing sites and a consensus processing site sequence is now emerging.     

Characterization and epitope mapping of human monoclonal antibodies to PDC-E2, the immunodominant autoantigen of primary biliary cirrhosis.

Further to define the epitopes of PDC-E2, the major autoantigen in primary biliary cirrhosis (PBC), we have developed and characterized five human monoclonal antibodies. These antibodies were derived by fusing a regional hepatic lymph node from a patient with PBC with the mouse human heterohybrid cell line F3B6. Previous studies of epitope mapping of PDC-E2 have relied on whole sera and have suggested that the immunodominant epitope lies within the inner lipoyl domain of the molecule. However, selective absorption studies using whole sera and a series of overlapping recombinant peptides of PDC-E2 have suggested that the epitope may also include a large conformational component. Moreover, several laboratories have suggested that autoantibodies against the 2-oxo acids dehydrogenase autoantigens are cross-reactive. The five monoclonal antibodies generated included three IgG2a and two IgM antibodies and were studied for antigen specificity using recombinant PDC-E2, recombinant BCKD-E2, histone, dsDNA, IgG (Fc), collagen and a recombinant irrelevant liver specific control, the F alloantigen. The antibodies were also used to probe blots of human, bovine, mouse and rat mitochondria. Finally, fine specificity was studied by selective ELISA and absorption against overlapping expressing fragments of PDC-E2. All five monoclonals, but none of the other mitochondrial autoantigens were specific for PDC-E2. In fact, although affinity purified antibodies to PDC-E2 from patients with PBC cross-reacted with protein X, the human monoclonals did not, suggesting that protein X contains an epitope distinct from that found on PDC-E2. Additionally, all three IgG2 monoclonals recognized distinct epitopes within the inner lipoyl domain of PDC-E2.     

HIV-1 gp41 by a common immunological epitope induces increased levels of antibodies against human interferon-β in HIV-1 positive individuals

Based on our finding that a similar epitope exists between human IFN-β (aa128–134) and HIV-1 gp41 (aa586–595), we examined 20 sera from healthy and 20 from HIV-1 infected individuals for IFN-β antibody levels by ELISA. The levels of anti-IFN-β antibody in sera from HIV-infected individuals were increased by about 160% in comparison with HIV-negative. We affinity-purified anti-gp41 antibodies from sera of HIV-1-infected individuals using rsgp41-sepharose column. One of three antibodies could recognize human IFN-β in comparison with antibodies from serum of a healthy individual. A mouse antiserum to human IFN-β recognized rsgp41 (recombinant soluble gp41, Env amino acid 539–684), while the normal mouse serum (pre-immune serum) did not bind to rspg41. These results indicate that a common immunological epitope exists between human IFN-β and HIV-1 gp41. The sequence-similarity suggests that this common immunological epitope may be located in the region aa128–134 of human IFN-β and the immunosuppressive domain (aa583–599) of HIV-1 gp41. The increased levels of antibodies against interferon-β in HIV-1 positive individuals may be explained by a common immunological epitope on human IFN-β and HIV-1 gp41.     

Therapy-induced antibodies to interferon-alpha 2a recognise its receptor-binding site.

Fifty-eight patients with chronic hepatitis B (HB) or C (HC) were treated with recombinant human interferon (rIFN)-alpha 2 and their sera were assayed for antibodies to rIFN-alpha 2c. Twelve of these patients produced low titres and two high titres of the antibodies. We localized the region which was recognised by the high-titre therapy-induced antibodies on the IFN molecule by testing the antibodies with a set of murine monoclonal antibodies (MoAbs) to IFN-alpha 2 in a competitive radioimmune assay (RIA). Only MoAbs with epitopes located in the amino-terminal portion of IFN-alpha 2 could inhibit the binding of radiolabelled IFN-alpha 2 by patients' sera. Our data indicate that the therapy-induced antibodies were directed to the receptor-binding domain of IFN-alpha 2 formed by amino acids (aa) 30-53. In accordance with this observation, human anti-IFN sera inhibited the binding of rIFN-alpha 2 to human cells.     

凡纳滨对虾主要过敏原-原肌球蛋白单克隆抗体的制备及其过敏原表位分析

目的制备凡纳滨对虾主要过敏原-原肌球蛋白的单克隆抗体(mAb),运用它们和虾过敏患者血清分析凡纳滨对虾原肌球蛋白的过敏原表位。方法纯化的凡纳滨对虾原肌球蛋白免疫Balb/c小鼠,间接ELISA和Western blot筛选并建立稳定分泌抗原肌球蛋白抗体的杂交瘤细胞株;腹水型mAbs经硫酸铵沉淀、Protein G亲和层析纯化;叠加ELISA分析mAbs的抗原结合表位;虾过敏患者血清lgE与mAbs的抑制Western blot和间接竞争ELISA分析原肌球蛋白的过敏原表位。结果共筛选出5株mAbs,它们之间叠加ELISA的叠加值均高于40%;其中B5和A5能显著抑制虾过敏患者血清IgE与原肌球蛋白的结合,抑制率分别为58.1%和48.6%,同时也能抑制66.7%和44.3%的血清IgE与虾蛋白粗提液反应。结论成功制备了5株分别结合原肌球蛋白不同抗原表位的单克隆抗体,其中B5和A5能结合其过敏原表位。     

Generation and characterization of human monoclonal neutralizing antibodies with distinct binding and sequence features against SARS coronavirus using XenoMouse

Passive therapy with neutralizing human monoclonal antibodies (mAbs) could be an effective therapy against severe acute respiratory syndrome coronavirus (SARS-CoV). Utilizing the human immunoglobulin transgenic mouse, XenoMouse, we produced fully human SARS-CoV spike (S) protein specific antibodies. Antibodies were examined for reactivity against a recombinant S1 protein, to which 200 antibodies reacted. Twenty-seven antibodies neutralized 200TCID(50) SARS-CoV (Urbani). Additionally, 57 neutralizing antibodies were found that are likely specific to S2. Mapping of the binding region was achieved with several S1 recombinant proteins. Most S1 reactive neutralizing mAbs bound to the RBD, aa 318-510. However, two S1 specific mAbs reacted with a domain upstream of the RBD between aa 12 and 261. Immunoglobulin gene sequence analyses suggested at least 8 different binding specificities. Unique human mAbs could be used as a cocktail that would simultaneously target several neutralizing epitopes and prevent emergence of escape mutants.     

Autoantibodies directed against the ribosomal P proteins are not only directed against a common epitope of the P0, P1 and P2 proteins.

The autoantibodies (aAbs) directed against the ribosomal P proteins (RPP aAbs) are known to react mainly against epitopes localized within the common C-terminal sequence of the three acidic ribosomal P proteins, P0, P1 and P2. In order to investigate the opportunity to select short recombinant peptides of this common C-terminal sequence to detect the RPP-aAbs, the location of the epitopes recognized by ribosomal proteins (RP) aAb(+)sera of systemic lupus erythematosus patients (SLE) was investigated. Immunoblotting and ELISA techniques using extracted or recombinant, entire or cleaved RPP showed that 55% of the RP aAbs were directed against the three ribosomal P0, P1, and P2 proteins. The epitopes recognized by the RPP aAbs are located not only within the C-terminal sequence common to the three proteins but also within the N-terminal sequence of the P2 or P1 protein. The other RP aAbs sera (45%) did not react with all three proteins but with some of them, and showed the following pattern: P0(+)P1(+); P1(+); P2(+); P0(+)and P1(+). They recognized epitopes located in the region of the C-terminal sequence of the protein but not common to the three proteins. In addition two out of the six monoclonal Abs produced by immunization of mice using the P1 protein did not react with the peptide N-65 or N-71 of the P2 protein or with the C-terminal sequence of the three proteins. In conclusion, this study showed that the RPP aAb in SLE patients are not only directed against epitopes within the C-terminal sequence shared by the three acidic ribosomal P proteins. In view of these data it seems necessary to be cautious in using only a C-terminal peptide of ribosomal P proteins in tests performed to detect RPP aAb in human sera.     

Mapping of B-cell epitopes of hemagglutinin protein of rinderpest virus

Monoclonal antibodies (mAbs) against secreted hemagglutinin (H) protein of rinderpest virus (RPV) expressed by a recombinant baculovirus were generated to characterize the antigenic sites on H protein and regions of functional significance. Three of the mAbs displayed hemagglutination inhibition activity and these mAbs were unable to neutralize virus infectivity. Western immunoblot analysis of overlapping deletion mutants indicated that three mAbs recognize antigenic regions at the extreme carboxy terminus (between amino acids 569 and 609) and the fourth mAb between amino acids 512 and 568. Using synthetic peptides, aa 569-577 and 575-583 were identified as the epitopes for E2G4 and D2F4, respectively. The epitopic domains of A12A9 and E2B6 mAbs were mapped to regions encompassing aa 527-554 and 588-609. Two epitopes spanning the extreme carboxy terminal region of aa 573 to 587 and 588 to 609 were shown to be immunodominant employing a competitive ELISA with polyclonal sera form vaccinated cattle. The D2F4 mAb which recognizes a unique epitope on RPV-H is not present on the closely related peste des petits ruminant virus HN protein and this mAb could serve as a tool in the seromonitoring program after rinderpest vaccination.     

抗rhNDPK-A单克隆抗体的制备及鉴定

目的 :研制抗rhNDPK A (recombinanthumannucleosidediphosphatekinase A)单克隆抗体 (mAb) ,并鉴定其特性。 方法 :以纯化的rhNDPK A免疫BALB/c小鼠 ,采用杂交瘤技术制备抗rhNDPK AmAb ;用免疫双扩散鉴定Ig亚类 ;West ernblot鉴定mAb的特异性 ;间接ELISA检测mAb的腹水效价、亲和常数 ,并进行表位分析。结果 :获得 6株可分泌特异性mAb的抗rhNDPK A的杂交瘤细胞系 2D9、8C7、13E2、15D9、15E3和 2 0D9,Ig亚类均为IgG1;其效价为 1× 10 -4～5× 10 -6;亲和常数为 4 .5× 10 -9～ 2 .8× 10 -10 mol/L ;共有3个抗原表位。结论 :获得抗rhNDPK A的mAb ,为进一步用于临床诊断和实验研究创造了条件     

骨桥蛋白单克隆抗体的制备及其特异性分析

为了研制骨桥蛋白(OPN)特异性单克隆抗体(mAb),并鉴定其特异性。本研究在毕赤酵母中表达具有良好免疫原性的重组人OPN蛋白的基础上,用重组人骨桥蛋白(rhOPN)免疫BALB/c小鼠,取其脾细胞与小鼠骨髓瘤细胞融合,间接ELISA筛选杂交瘤细胞,并结合免疫印迹对抗体的特异性进行鉴定,通过竞争抑制试验对单克隆抗体识别的抗原位点进行分析。结果共获得4株能够识别OPN不同抗原位点的mAb,亚类测定显示,3株为IgG1,1株为IgG2a。这些mAb能与重组人OPN特异性结合。本研究的四株抗体中,只有4G2B5能够检出与肿瘤转移密切相关的OPN-c的条带,预示该抗体可用于判断肿瘤预后和高转移肿瘤的临床检测。本研究成功获得了针对骨桥蛋白的特异性mAb,同时为进一步研究OPN蛋白的结构和功能提供了重要工具。     

以乙肝病毒核心蛋白为载体制备肠道病毒71 型非结构蛋白3D 单克隆抗体

目的：预测肠道病毒71型（EV71）非结构蛋白3D的表位,以HBc蛋白为载体展示多肽,制备并鉴定抗EV71-3D的特异性单克隆抗体（mAb）。方法：应用生物信息学方法分析预测出EV71-3D蛋白亲水性和免疫原性指数较高的多肽片段,并运用HBc颗粒型蛋白载体展示肽段,构建多肽融合蛋白,免疫BALB/c雌鼠,通过杂交瘤技术和亲和层析技术制备和纯化抗EV71-3D蛋白的特异性mAb,用间接ELISA、ELISPOT、IFA和IHC对mAb的性质进行初步鉴定。结果：构建表达分别嵌合3D蛋白34~43位氨基酸残基、 61~76位氨基酸残基、151~164位氨基酸残基的HBc重组蛋白,免疫并经过多轮克隆化筛选,获得抗EV71-3D单克隆抗体3E1,其亚类为IgG2a;免疫荧光试验、ELISPOT法和免疫组织化学染色结果显示其可与EV71特异性结合。结论：成功制备可特异性识别EV71的单克隆抗体3E1,为病毒的检测及进一步研究3D蛋白的功能奠定了基础,同时还验证了生物信息学技术与HBc颗粒型载体展示多肽技术相结合可快速高效地制备单克隆抗体。     

降钙素原抗原的表达及其抗体的制备与初步鉴定

目的:构建降钙素原(PCT)原核表达载体, 制备其多克隆抗体(pAb)和单克隆抗体(mAb), 并进行特性鉴定.方法:以甲状腺癌细胞cDNA文库为模板, 构建重组表达质粒pGEX- 4T-1-PCT和PET-32a-PCT, 在大肠杆菌中分别表达GST-PCT和His-PCT融合蛋白, 用His-PCT免疫家兔和BALB/c小鼠制备兔pAb和鼠mAb.通过ELISA法测定抗人PCT抗血清效价;用Western blot、 间接免疫荧光鉴定pAb的特异性.使用间接ELISA法筛选分泌特异性抗PCT的杂交瘤细胞株, 采用Western blot和间接免疫荧光鉴定mAb的特异性. 结果:构建了重组表达质粒pGEX- 4T-1-PCT和PET-32a-PCT, 并在大肠杆菌中表达、纯化.经ELISA法测得抗人PCT抗血清效价是1∶ 256 000.制备的抗PCT兔多克隆抗体可特异地识别重组和天然的PCT蛋白.获得8株可稳定分泌抗PCT的杂交瘤细胞株, 可识别重组人PCT蛋白, 其中有4株可特异性结合TT细胞质中的天然蛋白. 结论: 成功地制备出兔抗人PCT抗血清和8株抗PCT的mAb, 为进一步研究PCT在严重的细菌感染和脓毒症中的病理及生理作用奠定了基础.