新型血液代用品研究.pdf

M021 1. 2. 100029 100080; HSA Hb-HSA 132kD MALLS 1:1 90 30 60 Ringer 30 60 Hb-HSA 100 SE-HPLC 5 [1,2] αβ Baxer [8] Hemoglobin,Hb [3] [4] [5] [6] DBBF PEG [9] [10] [7] 157kD HSA Ringer Hb-HSA 90 30 60 60 1 1.1 Hb-HSA HSA 5 (3 [11] ) 2mg/ml, 20mmol/L - (pH5.7) 30:1 4 1h 1:2 HSA 4 9h DEAE-Sepharose Fast Flow 50mmol/L PBS (pH7.0) 5ml/min 0.1mol/LNaCl 50mM PBS (pH7.0) 280nm 405nm 5g/dl 4 1.2 Fiske ONE-TENTM Osmometer Fiske Associates, USA Evelyn 630nm 540nm [13] Hemox [12] ( TCS ) P50 Hill (SE-HPLC) Agilent 1100 TSK 3000SW 100mmol/L NaH2PO4/Na2HPO4 PBS 100mmol/L NaSO4 pH6.4 0.5ml/min MALLS DAWN EOS ,Wyatt Technology Co. USA HPLC 1.3 5 18~20g 0.5ml 5ml 2 270 280g Sprague-Dawley 300 350g Hb HSA [Hb]5g/dl Hb 5g/dl 3 102mmol/LNaCl, 4mmol/LKCl, 1.5mmol/L CaCl2, 28mmol/L 5 HSA ( Ringer pH7.4) 58ml/kg 30 6 0.045g/kg 0.3% BL-410E Ringer ,pH7.4 60 0.5mm 10min 14 0.5ml/min 3 10min 37 2 P0.05 xs 2 2.1 GPC 1 GPC MALLS 26min 148kD 22min 308kD 28.5min 75kD 33min 29min 308kD Hb HSA 132kD Hb-HSA 11 28.5min 32.5min Hb-HSA 1 1 89.7% Hb HSA 6.0 HSA Hb 3.5% mAU 1400 1200 1000 800 0.8% 157kD mAU Hb 180 150 120 HSA mAU 50 40 30 Hb-HSA conjugate 600 90 20 400 60 10 200 30 0 00 0 10 20 30 40 50 time/min time/min0 10 20 30 40 50 0 10 20 30 40 50 time/min 1 Fig. 1 Size-exclusion chromatography pattern of Hb-HSA conjugate Hemox P50 18.18mmHg Hill 2.10 OD OD OD P50 P50 P50 P50 P5016mmHg [14] P50 P5030mmHg [15] 1[5] [5] [16] 4.6~5.2 157kD [17] Table 1 Physicochemical properties of RBC, Hb-HSA conjugate and other Hb-based O2 carriers Hb-HSAconjugate Poly-Hb DBBF-Hb Blood (RBC) (nm) 30±5 47±17 7±2 Ca.8000 Mn (kDa) 148 510 72 Mw (kDa) 154 2154 66 P50 (mmHg) 18 20 32 28 3 Hill (cP) () () () (g/dl) (mmHg) 1.2 20 2.3 0.8 5 2.0 14.5 4.6~5.2 2.2 2.0 2.7-3.0 22 32 28 8.0 2.2 0.5 2.7 100 55 12-17 1.5 1.0 3-4 2.5 15.8 Ca.25 6.7~7.3 2.2 7 2.2.2 30 60 5 HSA Ringer 30 2 Hb-HSA Table 2 Change of blood pressure in the process of 30 exchange transfusion Groups Normal After bleeding After infusion 60min (0min) (10min