卵白リゾチーム - 愛媛大学gel gel matrix bead (b) solvent (c) (d) (e) small volume of...

卵白リゾチーム 0.2mm

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卵白リゾチーム

0.2mm

リゾチーム　（卵白）

PDB:1E8L (NMR)

Page 3: 卵白リゾチーム - 愛媛大学Gel Gel matrix bead (b) Solvent (c) (d) (e) Small Volume of effluent molecules Large molecules Gel beads Figure 5-7 Fundamentals of Biochemistry,

Page 4: 卵白リゾチーム - 愛媛大学Gel Gel matrix bead (b) Solvent (c) (d) (e) Small Volume of effluent molecules Large molecules Gel beads Figure 5-7 Fundamentals of Biochemistry,

Page 5: 卵白リゾチーム - 愛媛大学Gel Gel matrix bead (b) Solvent (c) (d) (e) Small Volume of effluent molecules Large molecules Gel beads Figure 5-7 Fundamentals of Biochemistry,

Page 6: 卵白リゾチーム - 愛媛大学Gel Gel matrix bead (b) Solvent (c) (d) (e) Small Volume of effluent molecules Large molecules Gel beads Figure 5-7 Fundamentals of Biochemistry,

①　ゲルろ過でBを除く②　陰イオン交換クロマトにAを吸着させ、CDを除いてから溶出する

※　クロマトの順序は逆でもよい。　ただし、陽イオン交換はだめ

解答例

3 0

２種

Page 7: 卵白リゾチーム - 愛媛大学Gel Gel matrix bead (b) Solvent (c) (d) (e) Small Volume of effluent molecules Large molecules Gel beads Figure 5-7 Fundamentals of Biochemistry,

[陽イオン交換はダメなワケ]

陽イオン交換A, B, C, D　　　　　 C (結合)

　　　　　　 A, B, D (素通り) 　　　　　A, D 　　　ゲルろ過

　　　陰イオン交換　　ゲルろ過A, B, C, D　　　　　 A, B (結合・溶出) A

　　C, D (素通り)

[正解例]

Page 8: 卵白リゾチーム - 愛媛大学Gel Gel matrix bead (b) Solvent (c) (d) (e) Small Volume of effluent molecules Large molecules Gel beads Figure 5-7 Fundamentals of Biochemistry,

アフィニティークロマトグラフィー–NADH cyt.b5 reductase–

SDS-PAGE

Page 9: 卵白リゾチーム - 愛媛大学Gel Gel matrix bead (b) Solvent (c) (d) (e) Small Volume of effluent molecules Large molecules Gel beads Figure 5-7 Fundamentals of Biochemistry,

基質

リガンド

樹脂

AMP-Sepharose chromatography

NADH

Page 10: 卵白リゾチーム - 愛媛大学Gel Gel matrix bead (b) Solvent (c) (d) (e) Small Volume of effluent molecules Large molecules Gel beads Figure 5-7 Fundamentals of Biochemistry,

Table 1Purification of NADH-cyt.b5 reductase from bovine brain

Page 11: 卵白リゾチーム - 愛媛大学Gel Gel matrix bead (b) Solvent (c) (d) (e) Small Volume of effluent molecules Large molecules Gel beads Figure 5-7 Fundamentals of Biochemistry,

限外ろ過器

遠心機で回転

Page 12: 卵白リゾチーム - 愛媛大学Gel Gel matrix bead (b) Solvent (c) (d) (e) Small Volume of effluent molecules Large molecules Gel beads Figure 5-7 Fundamentals of Biochemistry,

凍結乾燥機

減圧除去

氷　水蒸気

氷の昇華

エバポレーターとは違う！

水

冷却

Page 13: 卵白リゾチーム - 愛媛大学Gel Gel matrix bead (b) Solvent (c) (d) (e) Small Volume of effluent molecules Large molecules Gel beads Figure 5-7 Fundamentals of Biochemistry,

透析

Page 14: 卵白リゾチーム - 愛媛大学Gel Gel matrix bead (b) Solvent (c) (d) (e) Small Volume of effluent molecules Large molecules Gel beads Figure 5-7 Fundamentals of Biochemistry,

http://www.ach.ehime-u.ac.jp/biotec/

学内限定

講義ファイル

Page 15: 卵白リゾチーム - 愛媛大学Gel Gel matrix bead (b) Solvent (c) (d) (e) Small Volume of effluent molecules Large molecules Gel beads Figure 5-7 Fundamentals of Biochemistry,

Page 16: 卵白リゾチーム - 愛媛大学Gel Gel matrix bead (b) Solvent (c) (d) (e) Small Volume of effluent molecules Large molecules Gel beads Figure 5-7 Fundamentals of Biochemistry,

Page 17: 卵白リゾチーム - 愛媛大学Gel Gel matrix bead (b) Solvent (c) (d) (e) Small Volume of effluent molecules Large molecules Gel beads Figure 5-7 Fundamentals of Biochemistry,

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