title: Optimized method for preparation of IgG-binding bacterial magnetic nanoparticles.
abstract: In this study, the optimized method for designing IgG-binding magnetosomes based on integration of IgG-binding fusion proteins into magnetosome membrane in vitro is presented. Fusion proteins Mbb and Mistbb consisting of magnetosome membrane protein MamC and membrane associating protein Mistic from Bacillus subtilis as anchors and BB-domains of Staphylococcus aureus protein A as IgG-binding region were used. With Response Surface Methodology (RSM) the highest level of proteins integration into magnetosome membrane was achieved under the following parameters: pH 8.78, without adding NaCl and 55 s of vortexing for Mbb; pH 9.48, 323 mM NaCl and 55 s of vortexing for Mistbb. Modified magnetosomes with Mbb and Mistbb displayed on their surface demonstrated comparable levels of IgG-binding activity, suggesting that both proteins could be efficiently used as anchor molecules. We also demonstrated that such modified magnetosomes are stable in PBS buffer during at least two weeks. IgG-binding magnetosomes obtained by this approach could serve as a multifunctional platform for displaying various types of antibodies.
authors.affiliation: Faculty of Biology, Moscow State University, Moscow, Russia; Centre Bioengineering, Russian Academy of Sciences, Moscow, Russia.
authors.forename: Denis S
authors.initials: DS
authors.lastname: Grouzdev
authors.affiliation: Centre Bioengineering, Russian Academy of Sciences, Moscow, Russia.
authors.forename: Marina V
authors.initials: MV
authors.lastname: Dziuba
authors.affiliation: Centre Bioengineering, Russian Academy of Sciences, Moscow, Russia.
authors.forename: Denis V
authors.initials: DV
authors.lastname: Kurek
authors.affiliation: Centre Bioengineering, Russian Academy of Sciences, Moscow, Russia.
authors.forename: Alexander I
authors.initials: AI
authors.lastname: Ovchinnikov
authors.affiliation: Centre Bioengineering, Russian Academy of Sciences, Moscow, Russia.
authors.forename: Nadezhda A
authors.initials: NA
authors.lastname: Zhigalova
authors.affiliation: Centre Bioengineering, Russian Academy of Sciences, Moscow, Russia.
authors.forename: Boris B
authors.initials: BB
authors.lastname: Kuznetsov
authors.affiliation: Centre Bioengineering, Russian Academy of Sciences, Moscow, Russia.
authors.forename: Konstantin G
authors.initials: KG
authors.lastname: Skryabin
mesh_terms: D001426:Bacterial Proteins / Q000378:metabolism; D007074:Immunoglobulin G / Q000378:metabolism*; D058185:Magnetite Nanoparticles / Q000737:chemistry*; D055944:Magnetosomes / Q000737:chemistry*; D008565:Membrane Proteins / Q000378:metabolism; D011485:Protein Binding; D013205:Staphylococcal Protein A / Q000378:metabolism; D013211:Staphylococcus aureus / Q000378:metabolism
chemical_list: D001426:Bacterial Proteins; D007074:Immunoglobulin G; D058185:Magnetite Nanoparticles; D008565:Membrane Proteins; D013205:Staphylococcal Protein A