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

Table 1 Comparative quantification of protein bound along column performing

Condition of column	Initial protein (µg ± SD)^a	Flow-through (µg ± SD)^a	Bound protein (µg ± SD)^a	% of bound protein^b
Column set up
Recombinant LC3 binding control	50	12.67 ± 0.54	37.33 ± 0.54	74.65 ± 0.96
Empty pET17b competent binding control	50	25.58 ± 0.27	24.42 ± 0.27	49.24 ± 0.01
Bacterial protein (0.5 µg/µl) applying step
B. pseudomallei WT	36.54 ± 0.54	23.08 ± 1.63	13.46 ± 1.09	36.87 ± 3.53
B. pseudomallei rpoS mutant	37.12 ± 1.36	24.42 ± 0.27	12.69 ± 1.63	34.14 ± 3.15
Negative control^c	24.42 ± 0.27	19.42 ± 1.36	5.00 ± 1.09	20.50 ± 4.68

^aData was obtained from independent experimental replicate
^b% of bound protein was calculated from bound 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 wild type (WT) strain

ISSN: 2045-3701