title: Quantitative analysis of PMLA nanoconjugate components after backbone cleavage.
abstract: Multifunctional polymer nanoconjugates containing multiple components show great promise in cancer therapy, but in most cases complete analysis of each component is difficult. Polymalic acid (PMLA) based nanoconjugates have demonstrated successful brain and breast cancer treatment. They consist of multiple components including targeting antibodies, Morpholino antisense oligonucleotides (AONs), and endosome escape moieties. The component analysis of PMLA nanoconjugates is extremely difficult using conventional spectrometry and HPLC method. Taking advantage of the nature of polyester of PMLA, which can be cleaved by ammonium hydroxide, we describe a method to analyze the content of antibody and AON within nanoconjugates simultaneously using SEC-HPLC by selectively cleaving the PMLA backbone. The selected cleavage conditions only degrade PMLA without affecting the integrity and biological activity of the antibody. Although the amount of antibody could also be determined using the bicinchoninic acid (BCA) method, our selective cleavage method gives more reliable results and is more powerful. Our approach provides a new direction for the component analysis of polymer nanoconjugates and nanoparticles.
authors.affiliation: Department of Neurosurgery, Cedars-Sinai Medical Center, 110 N. 127 S. San Vincente, Advanced Health Science Pavilion A8220, Los Angeles, CA 90048, USA. hui.ding@cshs.org.
authors.forename: Hui
authors.initials: H
authors.lastname: Ding
authors.affiliation: Department of Neurosurgery, Cedars-Sinai Medical Center, 110 N. 127 S. San Vincente, Advanced Health Science Pavilion A8220, Los Angeles, CA 90048, USA. rameshwar.patil@cshs.org.
authors.forename: Rameshwar
authors.initials: R
authors.lastname: Patil
authors.affiliation: Department of Neurosurgery, Cedars-Sinai Medical Center, 110 N. 127 S. San Vincente, Advanced Health Science Pavilion A8220, Los Angeles, CA 90048, USA. jose.portilla@cshs.org.
authors.forename: Jose
authors.initials: J
authors.lastname: Portilla-Arias
authors.affiliation: Department of Neurosurgery, Cedars-Sinai Medical Center, 110 N. 127 S. San Vincente, Advanced Health Science Pavilion A8220, Los Angeles, CA 90048, USA. keith.black@cshs.org.
authors.forename: Keith L
authors.initials: KL
authors.lastname: Black
authors.affiliation: Department of Neurosurgery, Cedars-Sinai Medical Center, 110 N. 127 S. San Vincente, Advanced Health Science Pavilion A8220, Los Angeles, CA 90048, USA. julia.ljubimova@cshs.org.
authors.forename: Julia Y
authors.initials: JY
authors.lastname: Ljubimova
authors.affiliation: Department of Neurosurgery, Cedars-Sinai Medical Center, 110 N. 127 S. San Vincente, Advanced Health Science Pavilion A8220, Los Angeles, CA 90048, USA. eggehard.holler@cshs.org.
authors.forename: Eggehard
authors.initials: E
authors.lastname: Holler
mesh_terms: D000970:Antineoplastic Agents / Q000737:chemistry; D016503:Drug Delivery Systems; D007700:Kinetics; D008293:Malates / Q000737:chemistry*; D058726:Nanoconjugates / Q000737:chemistry*; D011108:Polymers / Q000737:chemistry*; D000068878:Trastuzumab / Q000737:chemistry
chemical_list: D000970:Antineoplastic Agents; D008293:Malates; D058726:Nanoconjugates; D011108:Polymers; C121060:poly(malic acid); D000068878:Trastuzumab