Purity and Recovery of crystallized mAb04c.

<p>*Protein redissolved in 100 mM sodium acetate pH 4.0.</p

Micrograph of crystals of mAb04c under polarized light.

<p>3 µl microbatch. Conditions: 1.5 µl 20 mg/ml mAb04c in 20 mM Tris, 50 mM Histidine, pH 7 plus 1.5 µl 12% (w/v) PEG 8000, 0.4 M calcium acetate in 0.2 M Imidazol, pH 7, RT. The broken crystal (intentionally) indicates that the dimension of crystal is about 100∼150 µm×10∼20 µm×10∼20 µm.</p

Micrograph of crystalline mAb04c crystallized from clarified culture supernatant.

<p>Crystallization condition: sitting drop. Reservoir: 0.1 M imidazol, 0.2 M calcium acetate, 9% w/v PEG 8000. 20 µL clarified culture supernatant (8.3 mg/ml mAb04c) plus 20 µl reservoir, RT. Scale bar 100 µm.</p

Phase diagram of mAb04c with PEG 8000 as precipitant.

<p>Buffer: 0.1 M imidazol, 0.2 M calcium acetate, pH 7.0, RT.</p

SDS-PAGE of HCP spiking experiment.

<p>Silver-staining. Lanes: (1) mAb04c, standard; (2) mother liquor with mAb04c and lysozyme; (3) washed mAb04c crystals from mother liquor with lysozyme; (4) the supernatant from the crystallization with lysozyme; (5) mother liquor with mAb04c and BSA; (6) washed mAb04c crystals from mother liquor with BSA; (7) the supernatant from the crystallization with BSA.</p

Coomassie blue stained none-reducing SDS-PAGE of mAb04c before and after crystallization or protein A purification.

<p>7 µg of IgG was loaded per lane. Lanes: (1) clarified mAb04c culture supernatant; (2) washed mAb04c crystals, redissolved in 100 mM sodium acetate pH 4.0; (3) mAb04c, purified via protein A chromatography.</p

SE-Chromatogram of the sample before and after crystallization.

<p>Elution buffer: 0.05 M Tris/0.15 M NaCl, pH 7; flow rate: 1 ml/min; wavelength: 225 nm. (A) Clarified mAb04c culture supernatant. Peak 3: mAb04c, 8.42 min; Peak 4: Contaminating Protein, 9.98 min; Peak 5: Histidine in buffer, 12.19 min. (B) washed mAb04c crystals, redissolved in 100 mM sodium acetate pH 4.0. Peak 2: mAb04c, 8.43 min.</p