• Title, Summary, Keyword: hemagglutination

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Development and Immunochemical Properties of Two Monoclonal Antibodies Specific to Human Chorionic Gonadotropin

  • Kim, You-Hee;Koh, Kwan-Sam
    • BMB Reports
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    • v.32 no.5
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    • pp.474-479
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    • 1999
  • Using a hybridoma technique, spleen cells of Balb/c mice immunized with human chorionic gonadotropin (hCG) were fused with NS-1 mouse myeloma cells. Two hybrid cell lines, clones KS-8 and KS-19, secreting monoclonal antibodies to hCG, were isolated. KS-8 and KS-19 belong to the immunoglobulin $G_1$ subclass. With the aid of a double-antibody radioimmunoassay, it was established that the KS-8 monoclonal antibody recognizes an immunodeterminant of the $\beta$-subunit of hCG, whereas the KS-19 monoclonal antibody recognizes an epitope present on the $\alpha$-subunit of hCG. The KS-8 monoclonal antibody specifically reacts with human chorionic gonadotropin and shows cross-reactivity of less than 0.3% to other related human glycoprotein hormones. On the other hand, using a hemagglutination test based on antibody-induced agglutination of sheep red blood cells coated with hCG, It was shown that only the KS-19 monoclonal antibody was capable of inducing a positive reaction, although both monoclonal antibodies had similar binding capacity to the coated cells. The results from the dual screening procedures demonstrate that KS-8 and KS-19 monoclonal antibodies show high sensitivity in two different assays, and are hence useful for the qualitative and quantitative determination of hCG by both radioimmunoassay and hemagglutination inhibition tests.

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Purification Efficiency of a Lectin from Maackia fauriei (솔비나무 유래 렉틴의 정제 효율)

  • Bae, Chan-Hyung;Kim, Ju-Cheol;Kim, Yu-Jeong;Kim, Sang-Gu;Na, Kwang-Heum;Park, Byung-Tae;Kim, Ha-Hyung
    • YAKHAK HOEJI
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    • v.51 no.4
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    • pp.259-263
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    • 2007
  • We previously reported the isolation of a sialic acid-specific lectin eluted from the bark of Maackia fauriei using alkaline buffer on a fetuin-affinity column. Application of a borate-based elution buffer in the present study increased the specific activity of purified lectin from crude protein extract by 2.6-fold, whilst only slightly decreasing the recovery by 1.13%. The biological properties of the lectin eluted with borate buffer were the same as those of the lectin eluted with alkaline buffer such as in terms of the hemagglutination activity, hemagglutination inhibition activity, molecular mass, purity, and cytotoxicity to human breast cancer cells. A prepared biotin-labeled lectin conjugate was used to investigate the binding to various glycoproteins. Our results indicate that eluting with borate buffer is more efficient than using alkaline buffer to isolate the lectin adsorbed in a fetuin-affinity column.

Comparative study on avian influenza virus antibody titer by hemagglutination inhibition test and enzyme-linked immunosorbent assay in the mass zone layer (산란계 밀집지역에서 혈구응집억제반응과 효소면역측정법을 이용한 가금인플루엔자 혈중항체가의 비교 조사)

  • 이정원;엄성심;이성재;서이원;서석열;정동석;송희종
    • Korean Journal of Veterinary Service
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    • v.27 no.1
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    • pp.81-87
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    • 2004
  • This study was conducted to investigate the similarity between hemagglutination inhibition (HI) test and enzyme-linked immunosorbent assay(ELISA), the HI titer and mean ratio S/P ratio) of avian influenza virus. To perform this study, the 1,457 sera of layers 21 farms in May, July and September, respectively. As a result of HI test, positive rates were 480 to 422 (92.1%) in May, 494 to 394(79.8%) in July and 483 to 402(83.2%) in September, and the mean antibody titer were 4.6, 4.3, 4.0 to 0.3 decreased, respectively. The positive rates by ELISA, 480 to 475(99.0%) in May, 494 to 485(98.2%) in July, 483 to 472(97.7%) in September, and the mean S/P ratio were 2.319, 2.557 and 2.380, respectively. The result of HI test and ELISA positive 480 to 422(92.1%), 475(99.0%), 494 to 394(79.8%), 485(98.2%) and 483 to 402(83.2%), 472(97.7%). Therefore, ELISA was shown more sensitive compare the HI titers.

In Vitro Antiviral Activity of Aqueous Extracts from Korean Medicinal Plants Against Influenza Virus Type A

  • Park, Kap-Joo;Lee, Hyung-Hoan
    • Journal of Microbiology and Biotechnology
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    • v.15 no.5
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    • pp.924-929
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    • 2005
  • Boiled-water extracts from 101 Korean medicinal plants were tested in vitro for their inhibitory activity against influenza virus type A by means of a modified hemagglutination inhibition test. Thirteen of the 101 extracts exhibited strong anti-influenza virus type A activity at concentrations of less than $780\;{\mu}g/ml$. Out of the above 13 extracts, MW-40 (Chaenomeles speciosa), MW-88 (Citrus junos), and MW-100 (Zingiber officinale) exhibited marked antiviral activity in the concentration range of $0.195\;{\mu}g/ml$ to 100 mg/ml, $0.0487\;{\mu}g/ml$ to 100 mg/ml, and $0.0487\;{\mu}g/ml$ to 100 mg/ml, respectively. The extracts MW-88 and MW-100 were not cytotoxic to red blood cells, whereas MW-40 showed very weak cytotoxicity in the concentration range of 50 mg/ml to 100 mg/ml. Therefore, the present results demonstrate that boiled water extracts of 2 Korean medicinal plants, MW-88 and MW-100, have strong anti-influenza virus type A activity and no cytotoxic effects, and they may inhibit attachment of the virus to the cell and may be used for prophylaxis.

Rabbit Hemorrhagic Disease Virus Variant Recombinant VP60 Protein Induces Protective Immunogenicity

  • Yang, Dong-Kun;Kim, Ha-Hyun;Nah, Jin-Ju;Song, Jae-Young
    • Journal of Microbiology and Biotechnology
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    • v.25 no.11
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    • pp.1960-1965
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    • 2015
  • Rabbit hemorrhagic disease virus (RHDV) is highly contagious and often causes fatal disease that affects both wild and domestic rabbits of the species Oryctolagus cuniculus. A highly pathogenic RHDV variant (RHDVa) has been circulation in the Korean rabbit population since 2007 and has a devastating effect on the rabbit industry in Korea. A highly pathogenic RHDVa was isolated from naturally infected rabbits, and the gene encoding the VP60 protein was cloned into a baculovirus transfer vector and expressed in insect cells. The hemagglutination titer of the Sf-9 cell lysate infected with recombinant VP60 baculovirus was 131,072 units/50 μl and of the supernatant 4,096 units/50 μl. Guinea pigs immunized twice intramuscularly with a trial inactivated RHDVa vaccine containing recombinant VP60 contained 2,152 hemagglutination inhibition (HI) geometric mean titers. The 8-week-old white rabbits inoculated with one vaccine dose were challenged with a lethal RHDVa 21 days later and showed 100% survival rates. The recombinant VP60 protein expressed in a baculovirus system induced high HI titers in guinea pigs and rendered complete protection, which led to the development of a novel inactivated RHDVa vaccine.

Purification and Characterization of the Lectins from Mushroom Flammulina velutipes (팽이버섯으로부터 Lectin의 정제와 특성)

  • Kim, Hyung-Suk;Son, Seung-Yeol;Hwang, Se-Young;Hong, Bum-Shik
    • Applied Biological Chemistry
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    • v.42 no.4
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    • pp.304-309
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    • 1999
  • Two Lectins, designated FVL-1 and FVL-2, were isolated and purified from the fruiting bodies of edible mushroom Flammulina veluripes using ammonium sulfate fractionation, ethanol treatment, DEAE-TOYPEARL ion-exchange column chromatography, and TSK-Gel HW-55F column chromatography. Specific activity increased 18 folds for FVL-1 and 7.9 folds for FVL-2 from ethanol treated sample. SDS-PAGE of FVL-1 and FVL-2 gave apparent molecular mass of 10.6 kDa and 37 kDa, respectively. FVL-2 agglutinated all type of human erythrocytes (A, B, AB, and O). However, FVL-1 agglutinated more human erythrocyte type O than type A, B, and AB. The hemagglutination activities of the FVL-1 were effectively inhibited by bovine submaxillary and porcine stomach mucins(BSM and PSM), fetuin, asialofetuin and cations, such as $Cu^{2+}$, $Mg^{2+}$, $Ca^{2+}$, $Mn^{2+}$ and $Fe^{2+}$. However, FVL-2 was not inhibited by any cations. The hemagglutination activities of the two lectins were not inhibited by the sugar, such as lactose, galactose and sugar derivatives. FVL-1 and FVL-2 were stable at pH $5{\sim}11$ and pH $4{\sim}7$, respectively. FVL-1 was stable below $55^{\circ}C$ and FVL-2 was below $45^{\circ}C$.

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Incidence of Antibodies for Toxoplasmosis of Cattle and Pigs in Jeonnam District (전남지역에서 사육하는 소와 돼지의 Toxoplasma 항체 조사성적)

  • Seo Doo-Seok;Kim Oh-Nam;Kang Byung-Kyu
    • Journal of the korean veterinary medical association
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    • v.16 no.1
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    • pp.29-34
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    • 1980
  • The latent infection of Toxoplasma in swine, Holstein cow and Korean native cattle raised in Jeonnam prefacture during 1978 was proved serologically. Of the 127 swine, 73($57.4\%$) cases were positive for the indirect hemagglutination test, and

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Hemagglutination of Tanned Chicken Erythrocytes by Newcastle Disease Virus ($B_1$ Strain) ($B_1$ 뉴캣슬병독에 의한 단닌산처리계적혈구의 혈구응집반응)

  • KIM SANG-YEOL
    • Journal of the korean veterinary medical association
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    • v.7 no.1
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    • pp.1-3
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    • 1963
  • Fazekas observed that fowl or human erythrocytes treated with a modifying dose of potassium pe-riodate remained agglutinable by influenza virus tut the adsorbed virus did not elute spontaneously. In this study, chicken erythrocytes treated with the high d

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Isolation and identification of canine adenovirus type 2 from a naturally infected dog in Korea

  • Yang, Dong-Kun;Kim, Ha-Hyun;Yoon, Soon-Seek;Lee, Hyunkyoung;Cho, In-Soo
    • Korean Journal of Veterinary Research
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    • v.58 no.4
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    • pp.177-182
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    • 2018
  • Canine adenovirus type 2 (CAV-2) infection results in significant respiratory illness in dogs. Isolating and culturing CAV-2 allows for investigations into its pathogenesis and the development of vaccines and diagnostic assays. In this study, we successfully isolated a virus from a naturally infected dog in Gyeonggi-do, Korea. The virus was propagated in Madin-Darby canine kidney (MDCK) and Vero cells and showed a specific cytopathic morphology that appeared similar to a bunch of grapes. The virus was first confirmed as CAV-2 based on these cytopathic effects, an immunofluorescence assay, hemagglutination assay, and electron microscopy. The viral titer of the isolate designated APQA1601 reached $10^{6.5}$ 50% tissue culture infections dose per mL in MDCK cells and exhibited no hemagglutination units with erythrocytes from guinea pig. The virus was also confirmed by polymerase chain reaction and next-generation sequencing. The APQA1601 strain had the highest similarity (~99.9%) with the Toronto A26/61 strain, which was isolated in Canada in 1976 when the nucleotide sequences of the full genome of the APQA1601 strain were compared with those of other CAV strains. Isolating CAV-2 will help elucidate the biological properties of CAV-2 circulating in Korean dogs.

Effects of Astragali Radix extract on the Humoral Immunotoxicity of Zinc Chloride (염화아연의 체액성 면역독성에 미치는 황기 추출물의 효과)

  • Chae, Byeong-Suk;Lim, Jong-Pil;Shin, Tae-Yong;Jeon, Hoon;Kim, Joung-Hoon
    • YAKHAK HOEJI
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    • v.44 no.1
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    • pp.1-8
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    • 2000
  • Effects of Astragali radix extract (AG) on the humoral immunotoxic responses of zinc chloride (Zn) were studied in ICR mice. Mice were divided into 4 groups (10 mice/group), and Zn was given to the mice, 1 hr after i.p. injection with 0.5 g/kg of AG, by i.p. injection daily for 10 days at a dose of 25 mg/kg. Mice were immunized and challenged with sheep red blood cells (SRBC). Zn treatment increased the relative weight of spleen compared with those in controls, but decreased the hemagglutination (HA) titer 2-mercaptoethanol-resistant HA (MER-HA) titer and splenic plaque forming cells (PFC). AG treatment significantly increased the relative weight of spleen, HA titer and PFC compared with those in controls. Combination of Zn and AG significantly increased the relative weight of spleen compared with those in controls, but decreased the HA titer, MER-HA titer and PFC. The relative weight of spleen was significantly increased in the Zn and AG combination group than those in Zn alone treatment group. But the HA titer, MER-HA titer and PFC were slightly increased in the group of combination. These results suggest that AG might slightly restore humoral immune responses lowered by an excess of zinc.

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