Production of a mouse hybridoma secreting monoclonal antibody highly specific to hemoglobin Bart's (γ4)

Hemoglobin (Hb) Bart's (γ4) was isolated and purified from Hb Bart's hydrops fetalis syndrome blood by CM-Sephadex C-50 chromatography. The isolated Hb Bart's was analyzed for its purity by high performance liquid chromatography. Balb/c mice were immunized intraperitoneally with Hb Ba...

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Main Authors: Makonkawkeyoon L., Pharephan S., Makonkowkeyoon S.
Format: Article
Language:English
Published: 2014
Online Access:http://www.scopus.com/inward/record.url?eid=2-s2.0-37849185172&partnerID=40&md5=5ea0497e73c0e11c13a541c1841e9421
http://cmuir.cmu.ac.th/handle/6653943832/1979
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Institution: Chiang Mai University
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spelling th-cmuir.6653943832-19792014-08-30T02:00:20Z Production of a mouse hybridoma secreting monoclonal antibody highly specific to hemoglobin Bart's (γ4) Makonkawkeyoon L. Pharephan S. Makonkowkeyoon S. Hemoglobin (Hb) Bart's (γ4) was isolated and purified from Hb Bart's hydrops fetalis syndrome blood by CM-Sephadex C-50 chromatography. The isolated Hb Bart's was analyzed for its purity by high performance liquid chromatography. Balb/c mice were immunized intraperitoneally with Hb Bart's. The immunized mouse splenic cells were hybridized with mouse myeloma, X63-Ag8.653, by polyethylene glycol. There were 12 hybridoma clones, out of several thousand culture wells, secreting antibody against purified Hb Bart's. However, when those 12 monoclonal antibodies (mAb) were tested with Hb Bart's (γ 4), HbF (α2γ2), HbH, HbE, and HbA2, there was only 1 hybridoma clone secreting mAb highly reactive to Hb Bart's with very low reactivity to HbF. A rabbit polyclonal antibody with relative high reactivity to Hb Bart's compared to HbF with the ratio of 2.4:1 was also produced by affinity column chromatography for the purpose of developing an enzyme-linked immunosorbent assay (ELISA) base for qualitative and quantitative determination of Hb Bart's in adult hemolysates. Preliminary results in quantitative determination of Hb Bart's in Hb solution of 3 α thalassemia families having at least 1 child with HbH disease and 6 normal subjects indicated that it was possible to quantify Hb Bart's by our developed ELISA with appropriate sensitivity and specificity. © 2006 Carden Jennings Publishing Co., Ltd. 2014-08-30T02:00:20Z 2014-08-30T02:00:20Z 2006 Article 10802924 10.1532/LH96.06027 17118769 LHAEA http://www.scopus.com/inward/record.url?eid=2-s2.0-37849185172&partnerID=40&md5=5ea0497e73c0e11c13a541c1841e9421 http://cmuir.cmu.ac.th/handle/6653943832/1979 English
institution Chiang Mai University
building Chiang Mai University Library
country Thailand
collection CMU Intellectual Repository
language English
description Hemoglobin (Hb) Bart's (γ4) was isolated and purified from Hb Bart's hydrops fetalis syndrome blood by CM-Sephadex C-50 chromatography. The isolated Hb Bart's was analyzed for its purity by high performance liquid chromatography. Balb/c mice were immunized intraperitoneally with Hb Bart's. The immunized mouse splenic cells were hybridized with mouse myeloma, X63-Ag8.653, by polyethylene glycol. There were 12 hybridoma clones, out of several thousand culture wells, secreting antibody against purified Hb Bart's. However, when those 12 monoclonal antibodies (mAb) were tested with Hb Bart's (γ 4), HbF (α2γ2), HbH, HbE, and HbA2, there was only 1 hybridoma clone secreting mAb highly reactive to Hb Bart's with very low reactivity to HbF. A rabbit polyclonal antibody with relative high reactivity to Hb Bart's compared to HbF with the ratio of 2.4:1 was also produced by affinity column chromatography for the purpose of developing an enzyme-linked immunosorbent assay (ELISA) base for qualitative and quantitative determination of Hb Bart's in adult hemolysates. Preliminary results in quantitative determination of Hb Bart's in Hb solution of 3 α thalassemia families having at least 1 child with HbH disease and 6 normal subjects indicated that it was possible to quantify Hb Bart's by our developed ELISA with appropriate sensitivity and specificity. © 2006 Carden Jennings Publishing Co., Ltd.
format Article
author Makonkawkeyoon L.
Pharephan S.
Makonkowkeyoon S.
spellingShingle Makonkawkeyoon L.
Pharephan S.
Makonkowkeyoon S.
Production of a mouse hybridoma secreting monoclonal antibody highly specific to hemoglobin Bart's (γ4)
author_facet Makonkawkeyoon L.
Pharephan S.
Makonkowkeyoon S.
author_sort Makonkawkeyoon L.
title Production of a mouse hybridoma secreting monoclonal antibody highly specific to hemoglobin Bart's (γ4)
title_short Production of a mouse hybridoma secreting monoclonal antibody highly specific to hemoglobin Bart's (γ4)
title_full Production of a mouse hybridoma secreting monoclonal antibody highly specific to hemoglobin Bart's (γ4)
title_fullStr Production of a mouse hybridoma secreting monoclonal antibody highly specific to hemoglobin Bart's (γ4)
title_full_unstemmed Production of a mouse hybridoma secreting monoclonal antibody highly specific to hemoglobin Bart's (γ4)
title_sort production of a mouse hybridoma secreting monoclonal antibody highly specific to hemoglobin bart's (γ4)
publishDate 2014
url http://www.scopus.com/inward/record.url?eid=2-s2.0-37849185172&partnerID=40&md5=5ea0497e73c0e11c13a541c1841e9421
http://cmuir.cmu.ac.th/handle/6653943832/1979
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