Active Structure Protection of Monoclonal Antibody Fusion Protein Etanercept†

doi:10.7503/cjcu20150695

Abstract

Abstract:

The thermodynamic stability of etanercept in different concentrations of the protective agent was investigated. The approaches of umbrella sampling and steered molecular dynamic simulation were adopted to study the dissociation of dimer etanercept with Gromacs software and amber99sb-ildn united atomic force field. The results indicated that the trehalose has ability to enhance the stability of dimer etanercept, and the protective effect of etanercept with trehalose and mannitol is better than with pure trehalose. In addition, the type of protective agent and alternative specific adsorption position can impact the effect of protecting active structure of etanercept observably.

Key words: Etanercept, Steered molecular dynamic, Umbrella sampling, Trehalose

CLC Number:

O629
Q341

TrendMD:

PAN Qi, LI Daixi, GUO Baisong, YANG Chunsheng, YANG Zhi. Active Structure Protection of Monoclonal Antibody Fusion Protein Etanercept^†[J]. Chem. J. Chinese Universities, 2016, 37(2): 274.

Figures/Tables 11

Fig.1 Sketch of monoclonal antibody fusion protein etanercept(A) and the structure of trehalose(B) and mannitol(C)

Table 1 Models for simulation and the detail components

Model	Etanercept/trehalose/mannitol	Number of water	Trehalose content(%)	Mannitol content(%)
Control	1/0/0	37481	0	0
THL-50	1/50/0	51156	1.73	0
THL-100	1/100/0	50383	3.45	0
THL-150	1/150/0	49604	5.16	0
THL-200	1/200/0	48829	6.86	0
THL-250	1/250/0	48041	8.54	0
THL-300	1/300/0	47244	10.23	0
THL-MAN	1/120/112	49068	4.12	2.06

Fig.2 Numbers of protection in different thickness of protective layer around atanercept in each system a-f respectively represents the etanercept solvation systems with 50, 100, 150, 200, 250 and 300 molecular numbers of trehalose. g-t and g-m respectively represents trehalose and mannitol in complex protection system.

Fig.3 Regional selective adsorption of protectant onto the surface of etanercept in different systems (A)—(F) represent the etanercept solvation systems with 50, 100, 150, 200, 250 and 300 molecular numbers of trehalose. (G) is trehalose and mannitol complex protection system. Closed electrostatic potential surface: etanercept; green: trehalose; cyan: mannitol. In order to show clearly, the molecular of water in systems is removed.

Fig.4 Curves of the pull force for dissociating etanercept dimer in different systems

Fig.5 Maximum pull force for dissociating etanercept dimer in different system s. THL-0; a. THL-50; b. THL-100; c. THL-150; d. THL-200; e. THL-250; f. THL-300; g. THL-MAN.

Fig.6 PMF free energy of etanercept dimer during dissociation in different systems

Table 2 Average PMF free energies and errors in etanercept dissociation in different systems

System	$E ?$ /(kJ·mol^-1)	Error/(kJ·mol^-1)	System	$E ?$ /(kJ·mol^-1)	Error/(kJ·mol^-1)
Control	314.46	6.32	THL-200	333.20	8.48
THL-50	487.63	8.63	THL-250	721.78	10.46
THL-100	684.51	8.20	THL-300	888.96	8.91
THL-150	721.60	9.77	THL-MAN	518.97	9.08

Table 2 Average PMF free energies and errors in etanercept dissociation in different systems

System	$E ?$ /(kJ·mol^-1)	Error/(kJ·mol^-1)	System	$E ?$ /(kJ·mol^-1)	Error/(kJ·mol^-1)
Control	314.46	6.32	THL-200	333.20	8.48
THL-50	487.63	8.63	THL-250	721.78	10.46
THL-100	684.51	8.20	THL-300	888.96	8.91
THL-150	721.60	9.77	THL-MAN	518.97	9.08

Fig.7 Adsorption of protectant within the range of 0.35 nm from etanercept dimer surface (A)—(F) represent the etanercept solvation systems with 50, 100, 150, 200, 250 and 300 molecular numbers of trehalose. (G) is trehalose and mannitol complex protection system. Dimer with scarlet and indigo: etanercept; green: trehalose; cyan: mannitol. In order to show clearly, the molecular of water in systems is removed.

Fig.8 Effect of trehalose adsorptive location on the structural stability of etanercept dimer (A) Adsorption of trehalose within the range of 0.35 nm from the etanercept dimer surface;(B) PMF free energy of etanercept dissociation in new system.

Fig.9 Effect of adsorptive location on the structural stability of etanercept dimer (A) Adsorption of protectant within the range of 0.35 nm from the etanercept dimer surface;(B) the PMF free energy of etanercept dissociation in new complex system.

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