• Proceedings of the Korean Society of Applied Pharmacology
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• 1994.04a
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• pp.269-269
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• 1994

GABA shunt에 관여하는 두 가지 효소인, GABA transminase와 Succinic semialdehyde reductase에 대한 단일 클론 항체를 생산하고 이들 항체의 특성을 살펴보았다. 소의 뇌에서 순수 정제된 효소를 동물에 주사한 후 immunodot-blot 분석법에 의하여 일차적으로 항체를 분비하는 hybridoma를 골라낸 후 생산된 단일 클론 항체가 특이적으로 이들 효소와 반응하는 가를 알아보기 위하여 Western blot 분석을 실시하였다. 뇌조직에서 추출한 총단백질을 SDS 전기영동법에 의하여 분리한 후 이들 항체를 처리한 결과, GABA-T에 대한 항체는 특이적으로 분자량이 50kDa에 해당하는 단백질 밴드만을 인지하였고 SSA reductase에 대한 항체의 경우 분자량이 34kDa 크기의 단백질 밴드와 반응하였다. 이들 분자량은 순수 정제된 소뇌의 효소 단백질의 크기에 해당하는 것을 확인하였다. 이들 소뇌의 효소에 대한 항체를 추적물체로 사용하여 다른 포유동물자 조류의 효소와 비교하는 cross-reactivity 연구를 수행하였다. 소, 돼지, 토끼, 쥐 (rat), 개, 고양이 그리고 닭의 뇌를 제거한 후 총단백질을 추출하고 Western blot을 해본 결과 GABA transaminase의 경우 조류를 제외하고 다른 포유동물에서는 같은 분자량의 단일 단백질 밴드를 확인하였고 SSA reductase의 경우 조류를 포함하는 모든 동물에서 같은 분자량의 밴드를 확인하였다 이상의 결과로 미루어 보아 포유류 뇌에 있는 이들 효소들은 변역학적으로 아주 유사하다고 사료된다.

• The Korean Journal of Zoology
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• v.32 no.4
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• pp.412-419
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• 1989

Monoclonal antibodies (MAbs) reacting with the plasmalemma of Amoeba proteus were produced. Specificity of the 3 MAbs was determined by transfer blotting of the SDS polvacryfamide gel. AMS antibody reacted with the mucopolysaccharide bands of the spacer gel, 220 KD and 50 KD proteins of the resolving gel. The maior glycoprotein bands (175 KD, 165 KD) and 50 KD protein of the plasmalemma were recognized by AUG antibody. A third, AMP antibody reacted with the 50 KD protein only. In immunofluorescence microscopy of the enzyme treated cells, the antigens of these MAbs were sensitive to proteases, but not sensitive to neuraminidase. In the assay of cell to substratum attachment after binding with the antibody, AMG and AMP antibodies exerted no effect, but AMS hindered the attachment and cell spreading. Thus the effective components of the plasmalemma in cell to substratum attachment appear to be the mucopolysaccharides and 220 KD protein. The membranes of latex particle infested phagosomes did not show any distinction from the plasmalemma in fluorescence microscopy. Phagosome membranes of amoebae appear to be derived from the plasma membrane without selection in terms of the antigen composition. Amoeba Proteus의 세포막과 반응하는 단세포군 항체를 생산하였다. SDS polyacrylamide gel을 transfer blotting하여 이들 항체의 반응 특이성을 조사해 본 결과 AMS 단항체는 PAS로 염색되는 spacer gel의 mucopolysaccharide 린드, resolving gel의 220 KD 및 50 KD 단백질과 반응하였으며, 세포막의 주요 당단백질인 175 KD 및 165 KD 빈드와 50 KD 단백질은 AMG 단항체에 의해서 인지되었다. 그리고 AMP단항체는 공통인 50 KD 단백질과 특이하게 반응하였다. 효소처리한 아메바의 면역형광칠미경적 조사에서 이들 항체에 대한 항원분자들은 모두 단백질분해효소에 민감하였으며 neuraminidase에 대해서는 변화가 없었다. 이들 항체를 결합시킨 아메바의 용기표면 부착 가능성을 분석한 결과 AMP 및 AMG 단항체는 아무런 영향을 미치지 못하였으며 AMS 단항체는 세포의 용기표면 부착 및 세포의 펴짐을 저해하였다. 따라서 아메바의 용기표면 부착은 mucopolysaccharide 및 220 KD 단백질에 의해서 매게되는 것으로 나타났다. 그리고 latex particle을 담고 있는 식포막은 면역형광형미경적 조사에서 세포막과 차이가 없었다. 따라서 겐포막은 항원 조성에 있어서 비 선택적으로 세포막에서 유도되는 것으로 나타났다.

• Korean Journal of Agricultural Science
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• v.14 no.2
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• pp.401-408
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• 1987

Nine monoclonal antibodies directed against infectious bovine rhinotracheitis virus (IBRV) were prepared by using cell hybridization technique, and the biological properties of the antibodies were investigated by means of immunofluorescence, serum neutralization, and electrophoretic analysis. Eight of 9 monoclonal antibodies reacted specifically with the antigenic constituents of IBRV, infectious laryngotracheitis virus, Marek's disease virus, turkey herpesvirus, hog cholera virus, porcine parvovirus and transmissible gastroenteritis virus. However, the remaining one, 26-2 clone, was found to be cross-reactive with pseudorabies virus. Two monoclonal antibodies, 7-C-2 and 12-A-2, which had neutralizing activity, were reactive with the molecular weights of 72 kilo daltons (72K) and 125K of IBRV proteins electrophoretically separated, respectively. The monoclonal antibody, 3-H-3, which is corresponding to 94K of IBRV proteins, revealed no neutralizing activity. The cross-reactive monoclonal antibody, 26-2, was proved by electrophoretical analysis to be reactive with 100K of IBRV proteins and 40K of pseudorabies virus.

• The Korean Journal of Zoology
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• v.31 no.4
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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.

• The Korean Journal of Zoology
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• v.39 no.4
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• pp.426-436
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• 1996

In previous studies, we have shown that lysosomal add phosphatase (LAP) activity increases at the dedifferentiation stage in the regenerating larval limbs of salamander, Hynobius leechii. Monoclonal antibodies against LAP were generated to determIne the spatial and temporal distribution of the protein In the regenerates.A total of 22 monoclonal antihodies recognizIng different epftopes of the protein were obtained, of which five strongly stained the regenerating limb by imunohistochemistry. in LAP immunohistochemical examination, LAP showed distribution coincident with the state of dedifferentiation, both spatially and temporally, in the limb regenerates. When unfractioned protein of regenerating salamander limbs were separated by gel electrophoresis and immunoblotted, the antibodies recognized a single protein band of 53 kl)a, which comigrates with a monomerlc subunit of IAR Using the anti-IAP antibodIes as probe, we investigated the cross-reactivities of LAPs from other sources. The immunoreadive bands on Western blots appeared to be the same In molecular mass-53 kl)a in axoloti and Xenopus, but no protein band was detected in mouse, Drosophila, or C. elegans.These results show that the antibodies generated in this study spedfically recognize Hynoblus leeclili IAp and that IAPs may be highiy conserved among amphibians. Furthermore, the distdbution of the protein is consistent with a role for LAP in the dedifferentiation process of limb regeneration.

• Microbiology and Biotechnology Letters
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• v.14 no.3
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• pp.219-223
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• 1986

In order to preparr the hybridoma cells which produce a monoclonal antibody to human fibroblast interferon(Hu IFN-$\beta$), spleen cells from BALB/cmice immunized with the purified Hu IFN-$\beta$ were fused with NS-O cells, a myeloma cell line. Forty hybrids with high titer among 1300 hybrids Isolated by an ELISA screening method were subcloned using the soft agarose cloning and limiting dilution methods, and 11 hybrids were selected. As a result of iso-typing the hybrids using the mouse monoclonal typing kit, two hybridomas were found to produce 1gG 2a type of monoclonal an-tibodies. The ascites fluid from nude mice inoculated intraperitoneally with the above hybridomas was removed and purified using a protein A-Sepharose CL-4B. Monoclonal antibody was proven to have only the heavy and light chains on SDS-polyacrylamide gel electrophoresis.

• Journal of the korean academy of Pediatric Dentistry
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• v.33 no.4
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• pp.587-596
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• 2006

Most of oral streptococci express the Antigen I/II (AgI/II) proteins, cell wall anchored adhesions. AgI/II protein binds to salivary agglutinin glycoprotein, a component of tooth pellicle and to ligands in other bacteria. These associations play important roles in bacterial colonization. Recently, it was reported that diverse host molecules also interact with AgI/II protein and that these interactions induce inflammatory responses from host cells. Among mutans streptococci containing -type hemolytic activity, Streptococcus mutans is a causative agent for dental caries. Compared with many other strains of S. mutans, GS-5 strain is unique in that this bacterium expresses truncated secretory AgI/II protein due to the nonsense mutation in the agI/II gene. This indicates that S. mutans GS-5 has a different clinical role and a recent report supported this idea based on the results from clinically isolated S. mutans strains. Previously, we had cloned agI/II gene from S. mutans GS-5 and generated recombinant N-terminal AgI/II protein. In this study, we further produced a hybridoma line expressing anti-AgI/II monoclonal antibodies named as 1C11A. This antibody showed high sensitivity to AgI/II protein in Western blot and ELISA. This new reagent will provide a basis for investigating the mechanisms of AgI/II-related diseases.

• The Science & Technology
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• v.16 no.5 s.168
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• pp.22-25
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• 1983

- 단일 클론항체, 의료계에 본격 진출 - 속성 양계 호르몬

• The Korea Swine Journal
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• v.21 no.11 s.243
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• pp.86-89
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• 1999

• Korean Journal of Microbiology
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• v.28 no.1
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• pp.6-12
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• 1990

The preS2 wequence of an adr hepatitis B virus was cloned and expressed in Escherichia coli as a $\beta$-galactosidase fusion polypeptide. Recombinant preS2 product interacted with the preS2-specific monoclonal antibody H8 which was induced by surface antigen particles isolated from a Korean gepatitis patient. The H8 showed only a minor cross-reactivity with recombinant preS2 product of adw2 subtype. Determination of nucleotide sequence of the adr preS2 revealed that twelve amino acid residue substitutions between adr and adw2 subtype sequences. The antigenic determinant to H8 must include some of these differences.