Monoclonal antibody interactions with micro- and nanoparticles: Adsorption, aggregation, and accelerated stress studies

Abstract

Therapeutic proteins are exposed to various wetted surfaces that could shed sub-visible particles. In this work we measured the adsorption of a monoclonal antibody (mAb) to various microparticles, characterized the adsorbed mAb secondary structure, and determined the reversibility of adsorption. We also developed and used a front-face fluorescence quenching method to determine that the mAb tertiary structure was near-native when adsorbed to glass, cellulose and silica. Initial adsorption to each of the materials tested was rapid. During incubation studies, exposure to the air-water interface was a significant cause of aggregation but acted independently of the effects of microparticles. Incubations with glass, cellulose, stainless steel or Fe 2 O 3 microparticles gave very different results. Cellulose preferentially adsorbed aggregates from solution. Glass and Fe 2 O 3 adsorbed the mAb but did not cause aggregation. Adsorption to stainless steel microparticles was irreversible, and caused appearance of soluble aggregates upon incubation. The secondary structure of mAb adsorbed to glass and cellulose was near-native. We suggest that the protocol described in this work could be a useful preformulation stress screening tool to determine the sensitivity of a therapeutic protein to exposure to common surfaces encountered during processing and storage.

Electrospray–differential mobility analysis of bionanoparticles

“…Experiments like those shown in Figure Other groups have reported that protein adsorption increases as a function of concentration at both stagnant to high shear flow conditions [106][107][108][109] which is in direct contrast to what is shown in Figure 4.3B. Nonetheless, the coverages of 3.5 -4.3 mg/m 2 determined here for concentrations ranging from 0.01 mg/mL to 0.1 mg/mL fall within the range of 2 -18 mg/m 2 reported in the literature for silica surfaces [106][107][108][109][110][111][112][113][114][115]. The reasons for the breadth of values may be attributed to differences in surface properties [116], ionic strength [106,107,117,118] , pH [113,114,117,119] and different immunoglobulins [113] .…”

Effects of shear on proteins in solution

“…during vial filling. Stainless-steel piston pumps are known to shed nanoparticles during operation that can lead to adsorption and subsequent aggregation of an IgG monoclonal antibody (Tyagi et al 2009;Bee et al 2009b) and indeed this may be a consequence of the micro-cavitation reported by van Reis and Zydney (2007). A key outcome of these studies was that particulates thought to provide substrates for aggregation in protein drug products were below the current particle size thresholds for which there is routine testing (e.g.…”

“…In this way the quality and safety of the commercial product might be ensured. Mahler et al (2009) describe some current techniques for detecting and characterizing aggregation, and some "emerging technologies", whilst Bee et al (2009b) suggest a protocol for assessing the sensitivity of therapeutic proteins to surfaces found in processing equipment and during storage. Table 1 shows a list of recommended screening studies that can be Table 1 here used to assess product sensitivity to shear forces and materials that are common in biopharmaceutical processes.…”

“…Bee et al (2009b) showed how a humanized IgG1 antistreptavidin interacts with such micro-and nano-particles. Adsorption to stainless steel microparticles was irreversible and led eventually to the formation of soluble protein aggregates.…”

Effects of Phosphate Buffer in Parenteral Drugs on Particle Formation from Glass Vials

“…[18][19][20][21] However, the Al-phosphate complex formation in other injectable drugs is unacceptable because these insoluble particles have the ability to harm the patient's veins. In particular, for development of antibody drugs, particles are a concern for biopharmaceutical manufacturers and regulatory officers 22) based on reports that particles in protein drugs may serve as nuclei triggering the aggregation of the protein, increasing the risk of immunogenicity, [23][24][25] and that particles may also enhance the immunogenicity of the protein drug by themselves. 26) Therefore, the phenomenon of particle formation from the storage of phosphate buffer solution in glass vials induced by interactions of the phosphate ions with Al eluted from the vial should be completely prevented in the field of injectable drug products.…”