| Japanese |
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| Title | 癌の免疫シンチグラフィにおける高免疫活性モノクローナル抗体の有用性に関する実験的研究 - (1) 高免疫活性抗体の分離・精製法の開発 - |
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| Subtitle | 原著 |
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| Authors | 横山邦彦* |
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| Authors(kana) | |
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| Organization | *金沢大学医学部核医学科 |
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| Journal | 核医学 |
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| Volume | 25 |
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| Number | 1 |
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| Page | 49-59 |
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| Year/Month | 1988/1 |
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| Article | 原著 |
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| Publisher | 日本核医学会 |
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| Abstract | 「要旨」放射性核種標識モノクローナル抗体による癌の診断および治療では, 用いる抗体の免疫活性(IR)が高く保持されていることが前提となる. 免疫活性の変化が抗体の体内動態に及ぼす影響を検討するための基礎的段階として, より高い免疫活性のモノクローナル抗体の分離・精製方法を開発し, その方法により得られた抗体を種々のin vitro cell binding assay (CBA)を用いて評価検討を行った. その結果既存の精製法で得られた悪性黒色腫に対する抗体96.5のF abフラグメントが, さらに2つの分画に, hydroxylapatite (HA)カラムを組み込んだ高速液体クロマトグラフィ(HPLC)によって分離された. CBAの結果2つの分画は, よりIRの低い抗体と高い抗体であることが示され, HA-HPLCによってIRの差に基づき抗体を効率よくかつ簡便に分離精製できることが明らかとなった. |
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| Practice | 臨床医学：一般 |
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| Keywords | Monoclonal antibody, F ab 96.5, Immunoreactivity, Affinity, Hydroxylapatite chromatography. |
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| English |
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| Title | Advantage of Highly Immunoreactive Monoclonal Antibodies in Radioimmunoscintigraphy for Tumor Detection : (I) New Purification System for Monoclonal Antibodies |
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| Subtitle | Original Articles |
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| Authors | Kunihiko YOKOYAMA |
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| Authors(kana) | |
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| Organization | Department of Nuclear Medicine, Kanazawa University School of Medicine |
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| Journal | The Japanese Journal of nuclear medicine |
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| Volume | 25 |
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| Number | 1 |
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| Page | 49-59 |
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| Year/Month | 1988/1 |
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| Article | Original article |
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| Publisher | THE JAPANESE SOCIETY OF NUCLEAR MEDICINE |
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| Abstract | [Summary] Immunoreactivity (IR) is the fraction of a monoclonal antibody (MoAb) preparation capable of binding to an excess of a specific antigen, One of the most important requirements for successful radioimmunoscintigraphy is to use a highly immunoreactive MoAb. To assess the effect of an antibody IR on biodistribution, a fast and simple purification method has been developed using a high performance liquid chromatography (HPLC) system equipped with a hydroxylapatite (HA) column. The column was eluted at ambient temperature with 0.12 M sodium phosphate buffer (pH 6.8). With this system, the F ab fragments from the MoAb 96.5 against the human melanoma associated p97 antigen were separated into two well-resolved peaks at retention times of 6 and 16 min. FEM-XII cells (human skin melanoma cell line) were used in a cell binding assay (CBA) to determine the maximal percent IR and the affinity constant of each HA-HPLC peak. The second peak from an 125I-F ab 96.5 showed approximately two times greater maximal binding than did the first peak, whereas the affinity constant for the two was the same. This indicated that the F ab 96.5 preparations used in this study were a mixture of more active and less active components. Moreover, prior to the HA-HPLC experiments, these preparations were analyzed with a gel filtration HPLC showing a single molecular weight peak. This suggested that the HA-HPLC separation was not based on molecular weight differences although the separation mechanism of HA has not yet been fully understood. Thereby, it is concluded that the HA-HPLC is a powerful tool to purify MoAbs into the higher immunoreactive fraction which has a potential advantage in tumor targeting. |
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| Practice | Clinical medicine |
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| Keywords | Monoclonal antibody, F ab 96.5, Immunoreactivity, Affinity, Hydroxylapatite chromatography. |
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