Investigation of host–pathogen interaction between Burkholderia pseudomallei and autophagy-related protein LC3 using hydrophobic chromatography-based technique

Table 2 Comparative quantification of eluted proteins obtained from each column condition

Condition of column	Bound protein (µg ± SD)^a	Flow-through (µg ± SD)^a	% of eluted protein^b
Elution step using 0.1 M glycine–HCl pH 2.7
B. pseudomallei WT	13.46 ± 1.09	8.30 ± 0.14	61.77 ± 2.89
B. pseudomallei rpoS mutant	12.69 ± 1.63	8.65 ± 0.11	68.56 ± 7.15
Negative control^c	5.00 ± 1.09	3.95 ± 0.11	89.82 ± 13.26
Washing step
B. pseudomallei WT	13.46 ± 1.09	4.60 ± 0.28	35.66 ± 2.10
B. pseudomallei rpoS mutant	12.69 ± 1.63	3.25 ± 0.18	26.59 ± 1.39
Negative control^c	5.00 ± 1.09	ND	ND

^aData was obtained from independent experimental replicate
^b% of eluted protein was calculated from eluted protein (μg) × 100/initial protein existed in column (μg)
^cNegative control represent to a column condition containing crude proteins without LC3 expression bait for protein of B. pseudomallei WT strain
ND represent to no determination because protein concentration could not be detected based on Bradford assay

ISSN: 2045-3701