Optimization of Unnicked β2-Glycoprotein I and High Avidity Anti-β2-Glycoprotein I Antibodies Isolation

Table 2

Isolation of human polyclonal high avidity IgG antibodies against β2-glycoprotein I: a representation of isolation efficiency calculation.

Starting material (plasma)	𝑉𝑜 (mL)	205
	^∗𝐶𝑜 HAv anti-𝛽2GPI (μg/mL)	33.9
	^∗𝑚𝑜 anti-𝛽2GPI (𝜇g)	6950
1. step	^∗𝑚start HAv anti-𝛽2GPI (𝜇g)	6950
IgG isolation	^∗𝑚isolated HAv anti-𝛽2GPI (𝜇g)	6049
(protein G column)	𝜂 (1. step)	0.870
2. step	^∗𝑚start HAv anti-𝛽2GPI (𝜇g)	6049
HAv a𝛽2GPI isolation	^∗𝑚isolated HAv anti-𝛽2GPI (𝜇g)	620
(𝛽2GPI column)	𝜂 (2. step)	0.102
	𝜂 total (%)	8.9%

Concentrations and calculated masses were determined by in-house anti-𝛽2GPI ELISA (*). 𝛽2GPI: 𝛽2-glycoprotein I; HAv anti-𝛽2GPI: high avidity anti-𝛽2GPI IgG antibodies; 𝑉_𝑜: volume of plasma used for isolation; 𝐶_𝑜: starting concentrations of HAv anti-𝛽2GPI; 𝑚_𝑜, 𝑚_start, and 𝑚_isolated: calculated masses of HAv anti-𝛽2GPI before isolation and before and after each step; and 𝜂: estimated efficiency of isolation.