Purification of Human Immunoglobulin G with Bathophenanthroline–Zn<sup>2+</sup>, –Fe<sup>2+</sup>, or –Cu<sup>2+</sup> Complexes
Background/Objectives: Pharmaceutical companies are aware of the ongoing effort to satisfy the increasing global demand for therapeutic-grade monoclonal antibodies (mAbs), an especially difficult challenge for poor and developing countries. We present a simple, economical, single-step purification a...
Saved in:
| Main Authors: | , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
MDPI AG
2025-05-01
|
| Series: | Antibodies |
| Subjects: | |
| Online Access: | https://www.mdpi.com/2073-4468/14/2/40 |
| Tags: |
Add Tag
No Tags, Be the first to tag this record!
|
| _version_ | 1849432146426462208 |
|---|---|
| author | Thisara Jayawickrama Withanage Ron Alcalay Olga Krichevsky Ellen Wachtel Ohad Mazor Guy Patchornik |
| author_facet | Thisara Jayawickrama Withanage Ron Alcalay Olga Krichevsky Ellen Wachtel Ohad Mazor Guy Patchornik |
| author_sort | Thisara Jayawickrama Withanage |
| collection | DOAJ |
| description | Background/Objectives: Pharmaceutical companies are aware of the ongoing effort to satisfy the increasing global demand for therapeutic-grade monoclonal antibodies (mAbs), an especially difficult challenge for poor and developing countries. We present a simple, economical, single-step purification approach at neutral pH for polyclonal human IgG (hIgG), which does not require any expensive ligands, chromatography columns, polymers, or membranes. Methods/Results: Instead, porous precipitates of commercial, recyclable aromatic [bathophenanthroline:cation] complexes were found to efficiently capture impurity proteins from CHO cells or <i>E. coli</i> lysate while maintaining the majority of the highly concentrated hIgG (5–15 mg/mL) in the supernatant. [(Batho)<sub>3</sub>:Zn<sup>2+</sup>] complexes were the most promising, resulting in hIgG with a purity of ≈95%, by SDS-PAGE. This purified hIgG is monomeric (by dynamic light scattering, DLS) and preserves the native secondary structure (by far UV circular dichroism spectroscopy, CD). The process yield is >90% (by densitometry) and is maintained after a 100-fold increase in the reaction volume, which required only proportional increases in reagents. Conclusions: Although Protein A chromatographic columns, the industry gold standard, have a limited binding capacity, are costly, and require familiarity with column maintenance, we are attempting, by our efforts, to help to produce a more efficient, simple, and economical purification platform. |
| format | Article |
| id | doaj-art-b422c8df4ccd477e91ec7a0df8d94b30 |
| institution | Kabale University |
| issn | 2073-4468 |
| language | English |
| publishDate | 2025-05-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Antibodies |
| spelling | doaj-art-b422c8df4ccd477e91ec7a0df8d94b302025-08-20T03:27:26ZengMDPI AGAntibodies2073-44682025-05-011424010.3390/antib14020040Purification of Human Immunoglobulin G with Bathophenanthroline–Zn<sup>2+</sup>, –Fe<sup>2+</sup>, or –Cu<sup>2+</sup> ComplexesThisara Jayawickrama Withanage0Ron Alcalay1Olga Krichevsky2Ellen Wachtel3Ohad Mazor4Guy Patchornik5Department of Chemical Sciences, Ariel University, Ariel 70400, IsraelIsrael Institute for Biological Research, Ness Ziona 7410001, IsraelDepartment of Chemical Sciences, Ariel University, Ariel 70400, IsraelFaculty of Chemistry, Weizmann Institute of Science, Rehovot 76100, IsraelIsrael Institute for Biological Research, Ness Ziona 7410001, IsraelDepartment of Chemical Sciences, Ariel University, Ariel 70400, IsraelBackground/Objectives: Pharmaceutical companies are aware of the ongoing effort to satisfy the increasing global demand for therapeutic-grade monoclonal antibodies (mAbs), an especially difficult challenge for poor and developing countries. We present a simple, economical, single-step purification approach at neutral pH for polyclonal human IgG (hIgG), which does not require any expensive ligands, chromatography columns, polymers, or membranes. Methods/Results: Instead, porous precipitates of commercial, recyclable aromatic [bathophenanthroline:cation] complexes were found to efficiently capture impurity proteins from CHO cells or <i>E. coli</i> lysate while maintaining the majority of the highly concentrated hIgG (5–15 mg/mL) in the supernatant. [(Batho)<sub>3</sub>:Zn<sup>2+</sup>] complexes were the most promising, resulting in hIgG with a purity of ≈95%, by SDS-PAGE. This purified hIgG is monomeric (by dynamic light scattering, DLS) and preserves the native secondary structure (by far UV circular dichroism spectroscopy, CD). The process yield is >90% (by densitometry) and is maintained after a 100-fold increase in the reaction volume, which required only proportional increases in reagents. Conclusions: Although Protein A chromatographic columns, the industry gold standard, have a limited binding capacity, are costly, and require familiarity with column maintenance, we are attempting, by our efforts, to help to produce a more efficient, simple, and economical purification platform.https://www.mdpi.com/2073-4468/14/2/40antibody purificationIgGnon-chromatographicligand-freeProtein A |
| spellingShingle | Thisara Jayawickrama Withanage Ron Alcalay Olga Krichevsky Ellen Wachtel Ohad Mazor Guy Patchornik Purification of Human Immunoglobulin G with Bathophenanthroline–Zn<sup>2+</sup>, –Fe<sup>2+</sup>, or –Cu<sup>2+</sup> Complexes Antibodies antibody purification IgG non-chromatographic ligand-free Protein A |
| title | Purification of Human Immunoglobulin G with Bathophenanthroline–Zn<sup>2+</sup>, –Fe<sup>2+</sup>, or –Cu<sup>2+</sup> Complexes |
| title_full | Purification of Human Immunoglobulin G with Bathophenanthroline–Zn<sup>2+</sup>, –Fe<sup>2+</sup>, or –Cu<sup>2+</sup> Complexes |
| title_fullStr | Purification of Human Immunoglobulin G with Bathophenanthroline–Zn<sup>2+</sup>, –Fe<sup>2+</sup>, or –Cu<sup>2+</sup> Complexes |
| title_full_unstemmed | Purification of Human Immunoglobulin G with Bathophenanthroline–Zn<sup>2+</sup>, –Fe<sup>2+</sup>, or –Cu<sup>2+</sup> Complexes |
| title_short | Purification of Human Immunoglobulin G with Bathophenanthroline–Zn<sup>2+</sup>, –Fe<sup>2+</sup>, or –Cu<sup>2+</sup> Complexes |
| title_sort | purification of human immunoglobulin g with bathophenanthroline zn sup 2 sup fe sup 2 sup or cu sup 2 sup complexes |
| topic | antibody purification IgG non-chromatographic ligand-free Protein A |
| url | https://www.mdpi.com/2073-4468/14/2/40 |
| work_keys_str_mv | AT thisarajayawickramawithanage purificationofhumanimmunoglobulingwithbathophenanthrolineznsup2supfesup2suporcusup2supcomplexes AT ronalcalay purificationofhumanimmunoglobulingwithbathophenanthrolineznsup2supfesup2suporcusup2supcomplexes AT olgakrichevsky purificationofhumanimmunoglobulingwithbathophenanthrolineznsup2supfesup2suporcusup2supcomplexes AT ellenwachtel purificationofhumanimmunoglobulingwithbathophenanthrolineznsup2supfesup2suporcusup2supcomplexes AT ohadmazor purificationofhumanimmunoglobulingwithbathophenanthrolineznsup2supfesup2suporcusup2supcomplexes AT guypatchornik purificationofhumanimmunoglobulingwithbathophenanthrolineznsup2supfesup2suporcusup2supcomplexes |