• 제목/요약/키워드: human antibodies

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계란 항체의 생산에 있어서 polymerization의 효과 (Effect of polymerization in inducing yolk antibodies)

  • 이경애
    • 한국생활과학회지
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    • 제2권1호
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    • pp.17-24
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    • 1993
  • Insulin (P)를 면역원으로 이용하여 insulin에 대한 균일한 항체집단의 대량유도 가능성 및 유도된 항체의 특성을 검토하였다. Insulin (P)에 의해 유도된 IgY는 insulin (M)에 의해 유도된 IgY 보다 조금 낮은 친화성을 나타내었으나 큰 차이는 없었다. 두 종류의 면역원에 의해 유도된 IgY는 대부분 insulin (M)을 인식하였으며, 같은 항원 특이성을 나타내었다.

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인터페론 알파에 대한 단세포 군항체의 제조 및 특성 (Production and Characteriuation of Monoclonal Antibodies against Human Interferon-$\alpha$)

  • Park, Kyung-Hee;Lee, Ihn-Sook
    • 한국동물학회지
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    • 제35권1호
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    • pp.1-7
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    • 1992
  • Seven monoclonal antibodies were produced by fusing splenocytes from Balb/C mouse immunized with partially purified human interferon-a (HUIFN-a) with NSO plasmacytoma cells. aery were identified as five IgG class (432.22: IgG2b/n, 460.52: IgG2b/a , 548.46: IgG2a/n , 573.10: IgG2b/h , 625.12: IgG2b/n ), one IgA class (460.50: IgA/n ) and one IsM class (465.27: IgA/n ), and all of them revealed highly sensitive to HUIFN- a IgG class monoclonal antibodies have pts ranged from 8.2 to 8.6. Ascites fluids produced from primed Balb/c mice and were purified through column chromatography. The cytopathic effect (CPE) inhibition assay to examine neutralization of HuIFU-a by IgG class monoclonal antibodies, gave that MAbs 460.52, 548.46, 573.10 can neutralize HUIFU- a arith varying degrees except 432.22. Therefore, it is deduced that these various monoclonal antibodies may recognize the distinct epitopes on HUIFN-a.

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Antibody Engineering for the Development of Therapeutic Antibodies

  • Kim, Sang Jick;Park, Youngwoo;Hong, Hyo Jeong
    • Molecules and Cells
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    • 제20권1호
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    • pp.17-29
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    • 2005
  • Therapeutic antibodies represent one of the fastest growing areas of the pharmaceutical industry. There are currently 19 monoclonal antibodies in the market that have been approved by the FDA and over 150 in clinical developments. Driven by innovation and technological developments, therapeutic antibodies are the second largest biopharmaceutical product category after vaccines. Antibodies have been engineered by a variety of methods to suit a particular therapeutic use. This review describes the structural and functional characteristics of antibody and the antibody engineering for the generation and optimization of therapeutic antibodies.

Expression Vectors for Human-mouse Chimeric Antibodies

  • Xiong, Hua;Ran, Yuliang;Xing, Jinliang;Yang, Xiangmin;Li, Yu;Chen, Zhinan
    • BMB Reports
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    • 제38권4호
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    • pp.414-419
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    • 2005
  • The production of recombinant antibodies has been generally recognized as time-consuming and labor-intensive. The aim of our study is to construct mammalian expression vectors containing the cDNA encoding the human constant regions and murine variable regions to massively and cost-effectively produce full-length chimeric antibodies. Unique restriction sites flanking the Ig variable region were designed to allow for the replacement of variable regions generated by PCR. Western blot analysis of the chimeric antibodies revealed that the expressed products were of the predicted size, structure and specificity. The usefulness of the vectors was confirmed by construction of human-mouse chimeric antibody-HCAb which secretes murine antibody against the human colorectal cancer. Selected in medium containing gradually increasing methotrexate (MTX), clones with increased expression of the product gene can be efficiently generated. The secretion of recombinant chimeric antibody-HCAb yielded $30\;pg\;cell^{-1}\;day^{-1}$ at $10^{-6}\;M$ MTX. With this high-level expression from pools, the convenient and rapid production of over 100 milligram amounts per liter of recombinant antibodies may be achieved, which indicates the significant roles of pYR-GCEVH and pYR-GCEVL in the production of chimeric antibodies.

사람 및 동물에 대한 소 엔테로바이러스 항체 분포 조사 (Existence of antibodies against bovine enterovirus in humans and various animals in Korea)

  • 박종현;김수미;방민우;이광녕;고영준;이향심;심항섭;조인수
    • 대한수의학회지
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    • 제49권3호
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    • pp.237-242
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    • 2009
  • Bovine enteroviruses (BEVs) were separated into two groups, BEV-1 and BEV-2. BEVs, found in cattle worldwide, usually cause asymptomatic infections and are excreted in the feces of infected animals. Antibodies against BEV have been found in different species including human, cattle, sheep, goats, dogs, horses and monkeys in the world. This study aimed to investigate prevalence of the neutralizing antibodies for BEVs in human and animals in Korea. Antibodies against BEV-1 in humans, cattle, pigs, goats, horses and dogs were shown to be 46.8%, 48.3%, 70.6%, 11.5%, 11.5% and 6.3% respectively. Also, antibodies against BEV-2 were shown to be 98.7%, 68.1%, 89.2%, 59.4%, 9.4% and 96.9% respectively. We found that the neutralizing antibodies against these viruses are common in Korea. The prevalences of antibodies against BEV-1 were lower than those against BEV-2 in humans and in all animals except horses. These results showed that the BEV is considered endemic in cattle in many regions in Korea.

Quantitation of Plasma Apolipoprotein A-I with a Sandwich Type Enzyme-Linked Immunosorbent Assay Using Monoclonal Antibodies

  • Lee, Min-Gyu;Kang, Jae-Seon;Jeong, Jae-Yeon;Jue, Dae-Myung;Kim, Hack-Joo
    • BMB Reports
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    • 제30권6호
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    • pp.390-396
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    • 1997
  • A sandwich-type enzyme-linked immunosorbent assay (ELISA) for the quantification of human apolipoprotein A-I (apoA-I) was developed using monoclonal antibodies. For this assay, we used three monoclonal antibodies to trap and detect apo A-I. HDAI16 and HDA15 monoclonal antibodies were used for trapping apoA-I and HDAI8 monoclonal antibody was for detecting apoA-I. These three monoclonal antibodies were produced by immunizing mice with high density lipoprotein (HDL) isolated from human plasma. By immunoblot analysis, these three monoclonal antibodies were specific to apoA-I and showed no cross-reactivities with other plasma proteins. The results of competition assays for epitope cross-reactivity test also verified that these monoclonal antibodies identified separate and distinct epitopes on HDL and apoA-I. Affinity constants of monoclonal antibodies were measured by ELISA. Their association constants ranged from $10^7$ to $10^8$ $M^{-1}$. For this assay, pure apoA-I was isolated by affinity chromatography using monoclonal antibodies. In this sandwich assay, the amount of HRP-labeled HDAI8 bound to apoA-I trapped by HDAI16 and HDAI5 was proportional to apoA-I concentration in the range of 0 to 500ng/ml. ApoA-I concentration in plasma was calculated from the linear regression equation of standard curve. The precision and reliability of the assays are reflected in the low intra-and interassay coefficients of variation that averaged 3.25% and 4.30%, respectively. This assay is sensitive, simple, reproducible, convenient in incubation interval, and does not use radioisotope: thus it can be widely applied in clinical laboratories.

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Tumor Marker 항원에 대한 단일 클론항체의 생성과 활용에 대한 연구. I. 태반형 Alkaline Phosphatase에 대한 모노클론항체의 생산과 분석 (Studies on the Generation and Application of Monoclonal Antibodies against Tumor Marker Antigen 1. Production and Characterization of Monoclonal Antibodies against Placental Alkaline Phosphatase)

  • 김한도;강호성
    • 한국동물학회지
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    • 제31권4호
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    • pp.300-308
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    • 1988
  • Human placental alkaline phosphatase (PLAP), one of the oncofetal antigen was purified from placentas through the procedures including butanol extraction, concanavalin A-Sephar-ose, DEAE-cellulose and Sephadex G-200 gel chromatography. Monoclonal antibodies (fibs) against human PMP were produced by hybridizing SP 210-Ag 14 mouse myeloma cells with spleen ceils of Balblc mice immunized with PLAP. Six stable monoclones uvere obtained by cloning tuvice in serial dilutions, and the monoclonal speclfidty of these MAbs was confirmed by biochemical and immunonogical criteria. Tumor marker의 하나인 태반형 alkaline phosphatase(PLAP)에 대한 단일 클론항체의 생산과 분석을 위하여, 태반조직을 재료로 butanol 추출법 및 concanavaline A-Sepharose, DEAE-cellulose, Sephadex G-200 gel 크로마토그라피법에 의하여 PLAP를 순수 분리하였다. 이를 항원으로 하여 하이브리도마 방법에 의해 항-PLAP 단일 클론항체를 생산 분비하는 안정된 6클론세포를 얻었으며 생화학적 및 면역학적 분석방법으로 이들의 단일 클론성을 확인하였다.

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Porphyromonas gingivali의 열충격단백-특이성 단클론항체의 개발 (Development of monoclonal antibody against Porphyromonas gingivalis heat shock protein)

  • 이니나;이주연;김성조;최점일
    • Journal of Periodontal and Implant Science
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    • 제37권1호
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    • pp.11-21
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    • 2007
  • Heat shock protein (HSP) is one of cellular protein commonly present in major periodontopathogenic bacteria as well as mammalian cells. The protein may play a role in the immunopathogenesis by modulating autoimmune reaction due to its high level of sequence homology between bacteria and human counterpart. Hence, identifying immunodomiant epitope of bacteria HSP that is cross-reactive to periodontopathogenic bacteria with a specificity to human HSP may comprise a critical strategy for development of a periodontal vaccine. The present study was performed to establish clones producing monoclonal antibody reactive to Porphyromonas gingivalis (p. gingivalis) HSP with a specificity to human HSP. 4 different hybridomas were cloned producing monoclonal IgG antibodies to P, gingivalis HSP and evaluated for their reactivity and specificity to other periodontopathogenic bacteria as well as to human HSP. These four monoclonal antibodies reacted with p. gingivalis HSP only with specificities to other bacteria tested and human HSP as well. The antigenic epitopes producing the 4 monoclonal antibody may be potentially developed as vaccine candidates. Further investigations are under way to identify more clones producing monoclonal antibodies reactive to P, gingivalis HSP and to other periodontopathogenic bacteria as well, while maintaining specificities to human counterpart.

Assessing the Archaeoparasitological Potential of Quids As a Source Material for Immunodiagnostic Analyses

  • Morrow, Johnica J.;Reinhard, Karl J.
    • Parasites, Hosts and Diseases
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    • 제54권5호
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    • pp.605-616
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    • 2016
  • In the present study, quids from La Cueva de los Muertos Chiquitos (CMC) were subjected to ELISA tests for 2 protozoan parasites, Toxoplasma gondii (n=45) and Trypanosoma cruzi (n=43). The people who occupied CMC, the Loma San Gabriel, lived throughout much of present-day Durango and Zacatecas in Mexico. The known pathoecology of these people puts them into at-risk categories for the transmission of T. gondii and T. cruzi. Human antibodies created in response to these 2 parasites can be detected in modern saliva using ELISA kits intended for use with human serum. For these reasons, quids were reconstituted and subjected to ELISA testing. All test wells yielded negative results. These results could be a factor of improper methods because there is no precedence for this work in the existing literature. The results could equally be a simple matter of parasite absence among those people who occupied CMC. A final consideration is the taphonomy of human antibodies and whether or not ELISA is a sufficient method for recovering antibodies from archaeological contexts. An additional ELISA test targeting secretory IgA (sIgA) was conducted to further examine the failure to detect parasite-induced antibodies from quids. Herein, the methods used for quid preparation and ELISA procedures are described so that they can be further developed by future researchers. The results are discussed in light of the potential future of quid analysis.

Synthetic Peptide-Based Enzyme-Linked Immunosorbent Assay for Human $\alpha$-Fetoprotein

  • Yoon, Mi-Chung;Lee, Hyun-Hee
    • 대한의생명과학회지
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    • 제7권3호
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    • pp.103-110
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    • 2001
  • $\alpha$-Fetoprotein(AFP) is a good marker for the detection of several diseases such as hepatocellular carcinoma, gonadal germ cell tumor, gastric tumor, and Down's syndrome. In this study, we developed ELISA, using synthetic peptides corresponding to the epitopes of AFP. Five kinds of peptides were synthesized from AFP to produce antibodies in rats that recognize AFP in human plasma as well as amniotic fluid and do not cross-react with serum albumin. All five kinds of antibodies showed good reactivities with their peptide-keyhole limpet hemocyanin conjugates. Anti-synthetic peptide 1 (R-N-E-Y-G-I-A-S-I-L, 4-13) antibody, in particular, reacted well with AEP as well as synthetic peptide 1-KLH but not with human serum albumin. The binding affinity(Kd) was 2.7$\times$10$^{-9}$M for peptide 1 and 6.8$\times$10$^{-8}$M for AEP. The range for measurement of AFP was 10~1,000 ng/ml. The within-assay and between-assay coefficients of variance(CV) were 4.83% and 10.97%, respectively. In a sample of 31 sera and 33 amniotic fluids, there was a good correlation between AFP values determined in this assay and those in a commercial kit. These results indicate that the antibodies against synthetic peptides corresponding to the epitopes of AFP are highly specific to APP and synthetic peptide-based ELISA would be useful for the measurement of human AFP.

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