Effect of Surfactants on Mechanical, Thermal, and Photostability of a Monoclonal Antibody

Abstract

The purpose of this work was to evaluate the effect of commonly used surfactants (at 0.01% w/v concentration) on mechanical, thermal, and photostability of a monoclonal antibody (MAb1) of IgG1 sub-class and to evaluate the minimum concentration of surfactant (Polysorbate 80) required in protecting MAb1 from mechanical stress. Surfactants evaluated were non-ionic surfactants, Polysorbate 80, Polysorbate 20, Pluronic F-68 (polyoxyethylene-polyoxypropylene block polymer), Brij 35 (polyoxyethylene lauryl ether), Triton X-100, and an anionic surfactant, Caprylic acid (1-Heptanecarboxylic acid). After evaluating effect of surfactants and determining stabilizing effect of Polysorbate 80 against mechanical stress without compromising thermal and photostability of MAb1, the minimum concentration of Polysorbate 80 required for mechanical stability was further examined. Polysorbate 80 concentration was varied from 0 to 0.02%. Mechanical stability was evaluated by agitation of MAb1 at 300 rotations per minute at room temperature for 72 h. Samples were analyzed for purity by SEC-HPLC, turbidity by absorbance at 350 nm, visible particles by visual inspection, and sub-visible particles by light obscuration technique on a particle analyzer. All non-ionic surfactants tested showed a similar effect in protecting against mechanical stress and did not exhibit any significant negative effect on thermal and photostability. However, Caprylic acid had a slightly negative effect on mechanical and photostability when compared to the non-ionic surfactants or sample without surfactant. This work demonstrated that polysorbate 80 is better than other surfactants tested and that a concentration of at least 0.005% (w/v) Polysorbate 80 is needed to protect MAb1 against mechanical stress.

What Makes Polysorbate Functional? Impact of Polysorbate 80 Grade and Quality on IgG Stability During Mechanical Stress

“…4 Surface active molecules, such as polysorbates (PSs), are common excipients in drug product formulations to increase colloidal stability and minimize adsorption of proteins to interfaces. 5,6 PSs are nonionic surfactants containing mainly sorbitan polyoxyethylene (POE) fatty acid esters. The protective role of PS in protein formulations has been thoroughly investigated, 7 and 2 main mechanisms have been elucidated: (1) PS acting as a chaperone for aggregation-prone hydrophobic sites on the protein surfaces (promoting the folded structure) 8,9 and (2) PS competing with proteins at interfaces (reducing interface exposure).…”

Manufacturing Challenges and Rational Formulation Development for AAV Viral Vectors

“…A rational formulation design chosen by optimizing solution conditions and the addition of excipients can significantly increase AAV stability and thus shelf-life. The usefulness of excipients on recombinant proteins has been extensively reported, 97,104,105,107,119,121,123,148 and excipients can in a similar way play a major role in the safety and efficacy of viral vectors. However, there is a lack of a rational, methodologically structured and systematic method of excipient selection.…”

Physicochemical Stability of Monoclonal Antibodies: A Review

“…Surfactants are generally added in mAbs formulations to reduce the exposure of hydrophobic regions and so decreasing proteinprotein interactions and interface-induced aggregation, also prevented by competition for adsorption sites. 129,130 Frequently used nonionic surfactants in mAb drug formulation are polysorbate 20 and polysorbate 80, 17,83 with poloxamers being a potential alternative. 131 Indeed, polysorbate 80 seems to be one of the most protecting surfactant against mechanical stresseinduced aggregation when compared to some other nonionic and anionic surfactants at the same concentration, 130 and also seems to be less perturbing toward mAb higher structure stability when compared to polysorbate 20.…”

“…132 This protection also depends on the protein to surfactant ratio. 29,93,130 In addition, surfactants have been shown to act as chemical chaperones, increasing rates of protein refolding and thus reducing aggregation. 46,47 For example, Gerhardt et al 133 reported that addition of polysorbate 20 to a solution containing various concentrations of a humanized IgG 1 antibody reduced particle formation during incubation or agitation with siliconized glass syringes, but noted a lack of correlation between the critical micelle concentration of the surfactant and its protective effect against aggregation.…”