A general platform for antibody purification utilizing engineered-micelles

Gunasekaran Dhandapani; Assaf Howard; Thien Van Truong; Thekke V. Baiju; Ellina Kesselman; Noga Friedman; Ellen Wachtel; Mordechai Sheves; Dganit Danino; Irishi N.N. Namboothiri; Guy Patchornik

doi:10.1080/19420862.2019.1565749

A general platform for antibody purification utilizing engineered-micelles

Gunasekaran Dhandapani, Assaf Howard, Thien Van Truong, Thekke V. Baiju, Ellina Kesselman, Noga Friedman, Ellen Wachtel, Mordechai Sheves, Dganit Danino, Irishi N.N. Namboothiri, Guy Patchornik^*

^*Corresponding author for this work

Research output: Contribution to journal › Article › peer-review

8 Scopus citations

Abstract

We introduce a new concept and potentially general platform for antibody (Ab) purification that does not rely on chromatography or specific ligands (e.g., Protein A); rather, it makes use of detergent aggregates capable of efficiently capturing Ab while rejecting hydrophilic impurities. Captured Ab are then extracted from the aggregates in pure form without co-extraction of hydrophobic impurities or aggregate dissolution. The aggregates studied consist of conjugated “Engineered-micelles” built from the nonionic detergent, Tween-20; bathophenanthroline, a hydrophobic metal chelator, and Fe ²⁺ ions. When tested in serum-free media with or without bovine serum albumin as additive, human or mouse IgGs were recovered with good overall yields (70–80%, by densitometry). Extraction of IgGs with 7 different buffers at pH 3.8 sheds light on possible interactions between captured Ab and their surrounding detergent matrix that lead to purity very similar to that obtained via Protein A or Protein G resins. Extracted Ab preserve their secondary structure, specificity and monomeric character as determined by circular dichroism, enzyme-linked immunosorbent assay and dynamic light scattering, respectively.

Original language	English
Pages (from-to)	583-592
Number of pages	10
Journal	mAbs
Volume	11
Issue number	3
DOIs	https://doi.org/10.1080/19420862.2019.1565749
State	Published - 3 Apr 2019
Externally published	Yes

Keywords

Antibody purification
Chromatography
IgG
Protein A
Protein G

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10.1080/19420862.2019.1565749

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title = "A general platform for antibody purification utilizing engineered-micelles",

abstract = " We introduce a new concept and potentially general platform for antibody (Ab) purification that does not rely on chromatography or specific ligands (e.g., Protein A); rather, it makes use of detergent aggregates capable of efficiently capturing Ab while rejecting hydrophilic impurities. Captured Ab are then extracted from the aggregates in pure form without co-extraction of hydrophobic impurities or aggregate dissolution. The aggregates studied consist of conjugated “Engineered-micelles” built from the nonionic detergent, Tween-20; bathophenanthroline, a hydrophobic metal chelator, and Fe 2+ ions. When tested in serum-free media with or without bovine serum albumin as additive, human or mouse IgGs were recovered with good overall yields (70–80%, by densitometry). Extraction of IgGs with 7 different buffers at pH 3.8 sheds light on possible interactions between captured Ab and their surrounding detergent matrix that lead to purity very similar to that obtained via Protein A or Protein G resins. Extracted Ab preserve their secondary structure, specificity and monomeric character as determined by circular dichroism, enzyme-linked immunosorbent assay and dynamic light scattering, respectively.",

keywords = "Antibody purification, Chromatography, IgG, Protein A, Protein G",

author = "Gunasekaran Dhandapani and Assaf Howard and Truong, {Thien Van} and Baiju, {Thekke V.} and Ellina Kesselman and Noga Friedman and Ellen Wachtel and Mordechai Sheves and Dganit Danino and Namboothiri, {Irishi N.N.} and Guy Patchornik",

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year = "2019",

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AU - Truong, Thien Van

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AU - Kesselman, Ellina

AU - Friedman, Noga

AU - Wachtel, Ellen

AU - Sheves, Mordechai

AU - Danino, Dganit

AU - Namboothiri, Irishi N.N.

AU - Patchornik, Guy

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N2 - We introduce a new concept and potentially general platform for antibody (Ab) purification that does not rely on chromatography or specific ligands (e.g., Protein A); rather, it makes use of detergent aggregates capable of efficiently capturing Ab while rejecting hydrophilic impurities. Captured Ab are then extracted from the aggregates in pure form without co-extraction of hydrophobic impurities or aggregate dissolution. The aggregates studied consist of conjugated “Engineered-micelles” built from the nonionic detergent, Tween-20; bathophenanthroline, a hydrophobic metal chelator, and Fe 2+ ions. When tested in serum-free media with or without bovine serum albumin as additive, human or mouse IgGs were recovered with good overall yields (70–80%, by densitometry). Extraction of IgGs with 7 different buffers at pH 3.8 sheds light on possible interactions between captured Ab and their surrounding detergent matrix that lead to purity very similar to that obtained via Protein A or Protein G resins. Extracted Ab preserve their secondary structure, specificity and monomeric character as determined by circular dichroism, enzyme-linked immunosorbent assay and dynamic light scattering, respectively.

AB - We introduce a new concept and potentially general platform for antibody (Ab) purification that does not rely on chromatography or specific ligands (e.g., Protein A); rather, it makes use of detergent aggregates capable of efficiently capturing Ab while rejecting hydrophilic impurities. Captured Ab are then extracted from the aggregates in pure form without co-extraction of hydrophobic impurities or aggregate dissolution. The aggregates studied consist of conjugated “Engineered-micelles” built from the nonionic detergent, Tween-20; bathophenanthroline, a hydrophobic metal chelator, and Fe 2+ ions. When tested in serum-free media with or without bovine serum albumin as additive, human or mouse IgGs were recovered with good overall yields (70–80%, by densitometry). Extraction of IgGs with 7 different buffers at pH 3.8 sheds light on possible interactions between captured Ab and their surrounding detergent matrix that lead to purity very similar to that obtained via Protein A or Protein G resins. Extracted Ab preserve their secondary structure, specificity and monomeric character as determined by circular dichroism, enzyme-linked immunosorbent assay and dynamic light scattering, respectively.

KW - Antibody purification

KW - Chromatography

KW - IgG

KW - Protein A

KW - Protein G

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A general platform for antibody purification utilizing engineered-micelles

Abstract

Keywords

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