Protein-poly(amino acid) precipitation stabilizes a therapeutic protein l-asparaginase against physicochemical stress

Wrap-and-Strip Technology of Protein–Polyelectrolyte Complex for Biomedical Application

“…Similar resistance was observed in the enzyme, L-asparaginase (Leunase for acute lymphoblastic leukemia; pI = 4.7), when in complexes with poly(lysine) (polyK; pKa = 10.5) of 4-5 kDa average molecular weight in 10 mM MOPS buffer (pH 7.0) [34]. L-Asparaginase-polyK precipitates also showed higher resistance to thermal inactivation, agitation, and oxidation.…”

“…The residual immunoreactivities of native adalimumab were 20.7% (0.5 mg/ml antibody) and 53.8% (5.0 mg/ml) following agitation, while those of polyE complex precipitation remained at approximately 100%. More importantly, the protein aggregates as part of the polyE complex were smaller in size than those without polyE following exposure to agitation stress [34].…”

“…Thus, the driving force behind protein-poly(amino acid) complex formation is mainly electrostatic interaction. Note that the resolubilized-state protein displays its original function and structure, as is the case with various kinds of proteins, such as the therapeutic monoclonal antibodies of Panitumumab, Etanercept, Adalimumab, Infliximab, Omalizumab, and Rituximab; the peptide hormone of Carperitide; the enzyme drug of L-asparaginase; and the globins of thyroglobulin and immunoglobulin G [32][33][34]. The recovery of activity and secondary structure implies that the process of PPC precipitation does not affect the tertiary structure of protein.…”

Aggregative protein–polyelectrolyte complex for high-concentration formulation of protein drugs

“…We have reported that the aggregative PPC protected unfavorable inactivation and aggregation of therapeutic proteins from the agitation stresses [20,21]. The stabilization effects of aggregative PPC have been tested as follow: (i) suspension (step 2 in Figure 2) and precipitation (step 3 in Figure 2) of therapeutic proteins and poly(amino acid) complexes were prepared; (ii) the mechanical stress including thermal and shaking were loaded on the sample; (iii) a salt solution (final concentration of 150 mM) was added into the sample; and then (iv) the residual activity and protein concentration of the redissolved proteins were determined.…”

“…Despite the disadvantage of aggregation state protein, we have proposed a unique approach to the concentration of protein solutions using protein aggregation with polyelectrolytes [18][19][20][21].…”