Heparin removal from human plasma using molecular imprinted cryogels

Figures & data

Table I. The optimization of the HpMIP columns.

Columns	HpMIP1	HpMIP2	HpMIP3	HpMIP4	HpMIP5	HpMIP6	HpMIP7	HpMIP8
HEMA/MBAAm (mol/mol)	4.0	6.0	8.0	6.0	6.0	6.0	6.0	6.0
Hp/DMAPM (mol/mol)	1/500	1/500	1/500	1/500	1/750	1/1000	1/500	1/500
HEMA (mol)	8.29	10.7	9.36	10.7	10.7	10.7	10.7	10.7
Hp (mol)	0.75	0.75	0.75	0.75	0.75	0.75	0.75	0.75
mg Hp/g MIP	8	8	8	8	8	20	40	60
Complex amount (× mol)	× 1	× 1	× 1	× 1	× 1	× 2.5	× 5.0	× 7.5

Figure 1. SEM photos of HpMIP (A, B) and NIP (C, D) columns.

Figure 2. FTIR spectrum of Hp, DMAPM and Hp–DMAPM complex (A), HpMIP (bottom) and NIP (upper) column (B).

Table II. Coagulation times of control, HpMIP and NIP cryogels.

	Control	HpMIP	NIP
PT (s)	29	32	30
APTT (s)	87.43	117	106.5

Figure 3. (A) Maximum Hp adsorption capacities of HpMIP2, HpMIP6, HpMIP7 and HpMIP8 columns. (B) Effect of initial Hp concentration on adsorption capacities of HpMIP, NIP and control (PHEMA) column. m_dry: 0.284 g; V: 5.0 mL; Flow rate 1.0 mL/min, t: 25°C.

Table III. Langmuir and Freundlich adsorption constants and correlation coefficients for Hp.

Experimental Qexp (mg/g)	Langmuir constants			Freundlich constants
	Qmax (mg/g)	b (g/mL)	R^2	Qf (mg/g)	n	R^2
9.2	11.9	17.6	0.95	6.12	0.64	0.89

Figure 4. Chromatogram of Hp separation with (A) HpMIP column; (B) NIP column from binary Hp:CHND, Hp:HSA, Hp:IgG solutions. (C) Chromatograms of Hp removal from Hp-loaded human blood plasma. (D) Back pressure property of HpMIP column.

Table IV. Selectivity coefficients of the HpMIP column.

	Sample (mol:mol)		t(min)	k’	α	IF	SF	N
HpMIP cryogel column	Hp:HSA 1000:100	Hp	14.61	53.11	1.55	1.0	1.087	15327
		has	9.5	34.18		0.98		672
	Hp:IgG 1000:100	Hp	14.58	53.0	1.56	1.0	1.089	17347
		IgG	9.44	33.96		0.97		828
	Hp:CHND 1000:500	Hp	14.56	52.93	1.56	1.0	1.089	16592
		CHND	9.43	33.92		0.97		526

Table V. Selectivity coefficients of the NIP columns.

	Sample (mol:mol)		t(min)	k’	α	N
NIP Cryogel column	Hp:HSA 1000:100	Hp	14.02	50.9	1.43	4700
		HSA	9.89	35.63		1572
	Hp:IgG 1000:100	Hp	14.05	51.03	1.43	2734
		IgG	9.89	35.62		1712
	Hp:CHND 1000:500	Hp	14.09	51.18	1.42	3507
		CHND	9.99	36.0		1629

Table VI. Removal amounts of Hp from human plasma for HpMIP, NIP and plain PHEMA cryogel columns.

Columns Plasma samples	HpMIP		NIP		Plain PHEMA
	Removed Hp amount		Removed Hp amount		Removed Hp amount
	U/mL	%	U/mL	%	U/mL	%
28 U/mL	25.2	90	23.8	85	0.98	3.5
10 U/mL	8.7	87	8	80	0.27	2.7