TOSOH
CUSTOMER MAGAZINE
The impact of chaotropic agents, such as guanidinium hydrochloride and
urea on static binding capacity of TOYOPEARL AF-rProtein A HC-650F has
been investigated. Aliquots of the resin have been incubated for 70 hours
in solutions of these agents at different concentrations. Static binding ca-
pacities are shown in Figure 3.
Thermo-treated resin and resin that has been incubated with chaotropic
agents were packed into 6.6 mm ID x 5 cm L columns. MAb feedstock
was purified with thermo-treated resin and the protein A elution pool was
analyzed with regards to their host cell protein (HCP) content. HCP titers
were determined using CHO HCP ELISA kit 3G (Cygnus Technologies).
TOYOPEARL AF-rProtein A HC-650F resin treated with sodium phosphate
buffer at room temperature served as a reference. Log reduction values
are roughly 2.9 for all of the tested conditions. No significant decrease in
HCP removal after thermo-treatment or incubation with chaotropic agents
is observed (Figure 4).
CIP based on thermo-treatment at 90 °C could be a valuable alternative
in case commonly used CIP strategies do not fully prevent form bacterial
growth. This seems particularly applicable to preparative scale in a stain-
less steel environment, since lab-scale applications often use temperature
sensitive hardware. Further cycling experiments will be employed to shed
light on the resin life time after exposure to periodic thermo-treatment.
AUTHORS:
JUDITH VAJDA, ELKE PROHASKA, MAIK RÖHL, EGBERT MÜLLER,
TOSOH BIOSCIENCE GMBH
[1] ECFR — CODE OF FEDERAL REGULATIONS
, (N.D.).
/
CGI-BIN/TEXT-IDX?SID=19AA3B822B3B03F66FF441B4D566452F&MC=TRUE&NODE=
SE21.2.129_180&RGN=DIV8 (ACCESSED MARCH 10, 2016).
[2] GENERAL CHAPTERS:
<1231> WATER FOR PHARMACEUTICAL PURPOSES, (N.D.).
-
COPEIA.CN/V29240/USP29NF24S0_C1231.HTML (ACCESSED MARCH 10, 2016).
[3] WHO GOOD MANUFACTURING PRACTICES:
WATER FOR PHARMACEUTICAL
USE, N.D.
/
TRS_970_ANNEX2.PDF.
FIGURE 3 :
STATIC BINDING CAPACITY OF TOYOPEARL AF-rProtein A HC-650F
AFTER 70 HOURS INCUBATION IN VARIOUS DIFFERENT CHAOTROPIC SOLUTIONS.
INCUBATION IN 2 M UREA INCREASES STATIC BINDING CAPACITY. GUANIDINIUM
HYDROCHLORIDE OR 100 % ETHANOL HAVE MINOR IMPACT ON STATIC BINDING
CAPACITY OF THE RESIN.
100 % ethanol
2 M gua/HCI
1 M gua/HCI
4 M urea
2 M urea
reference
160
140
120
100
80
60
40
20
0
static binding capacity [%]
FIGURE 4 :
LOG REDUCTION VALUES FOR HOST CELL PROTEIN REMOVAL. THE RE-
FERENCE SAMPLE HAS BEEN TREATED WITH SODIUM PHOSPHATE BUFFER AT ROOM
TEMPERATURE. NEITHER THERMO-TREATMENT NOR INCUBATION OF THE RESIN FOR
70 HOURS IN DIFFERENT CONCENTRATIONS OF UREA OR GUANIDINIUM HYDROCHLO-
RIDE OR 100 % ETHANOL SIGNIFICANTLY AFFECTS HOST CELL PROTEIN REMOVAL.
100 % EtOH
2 M gua/HCI
1 M gua/HCI
4 M urea
2 M urea
90 °C
reference
3.0
2.5
2.0
1.5
1.0
0.5
HCP Log reduction value
