One single method to produce native and Tat-fused recombinant human α-synuclein in Escherichia coli

BMC Biotechnology

Table 1 Purification of native AS and TAT-AS from E. coli starting from 1 L culture (≈ 2.5 g cell paste)

Purification step	Total protein (mg)	AS (mg)	AS purity ^a(%)	Yield ^b(%)
Whole cell extract ^c	210 [185] ^d	35 [22.2]	16.7 [12.0]	100 [100]
Osmotic shock	40 [29] ^d	30 [20]	75 [69]	85 [90.1]
HiTrap chromatography	16 [11.8] ^e	16 [11.8]	> 95 [> 95]	53 [59]

The value obtained for the purification of TAT-AS is shown in brackets.
^a The purity of AS at each step of purification was estimated by SDS–PAGE.
^b Ratio of the amount of AS obtained at each step to the total amount of AS estimated in the whole cell extract.
^c Supernatant of the cell lysate by sonication.
^d Determined by Bradford assay with bovine serum albumin as a standard.
^e Determined spectrophotometrically.

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