Magnetic hydrophobic-charge induction adsorbents for the recovery of immunoglobulins from antiserum feedstocks by high-gradient magnetic fishing

Cláudia Sg Gomes, Adedayo Fashina, Alfred Fernández-castané, Timothy W Overton, Timothy J Hobley, Eirini Theodosiou, Owen Rt Thomas

Research output: Contribution to journalArticle

Abstract

BACKGROUND The extraction of biopharmaceuticals from plasma and serum often employs overly complicated antiquated procedures, that can inflict serious damage on especially prone protein targets and which afford low purification power and overall yields. Here we describe systematic development of a high-gradient magnetic fishing process for recovery of immunoglobulins from unclarified antiserum. RESULTS Non-porous superparamagnetic particles were transformed into hydrophobic-charge induction adsorbents and then used to recover immunoglobulins from rabbit antiserum feedstocks. Comprehensive characterisation tests conducted with variously diluted clarified antiserum on a magnetic rack revealed that immunoglobulin binding was rapid (equilibrium reached in <45 s), strong (Kd <0.1 mg mL-1), of high capacity (Qmax = 214 mg g-1), and pH and ionic strength dependent. In a high-gradient magnetic fishing process conducted with the same adsorbent, and a conventional ‘magnetic filter + recycle loop’ arrangement, >72% of the immunoglobulin present in an unclarified antiserum feed was recovered in 0.5 h in >3-fold purified form. CONCLUSIONS Fast magnetic particle based capture of antibodies from an unclarified high-titre feed has been demonstrated. Efficient product recovery from ultra-high titre bioprocess liquors by high-gradient magnetic fishing requires that improved magnetic adsorbents displaying high selectivity, ultra-high capacity and operational robustness are used with 'state-of-the-art’ rotor-stator magnetic separators.
Original languageEnglish
JournalJournal of Chemical Technology and Biotechnology
Early online date2 Feb 2018
DOIs
Publication statusPublished - 2 Feb 2018

Fingerprint

Adsorbents
Feedstocks
Immunoglobulins
Immune Sera
fishing
Magnetic separators
Recovery
Antibodies
Stators
antibody
Purification
purification
serum
Rotors
fold
Proteins
Plasmas
plasma
damage
protein

Bibliographical note

Funding: Biotechnology & Biological Sciences Research Council (BBSRC) and the Danish Council forS trategic Research( DetStrategiskeForskningsråd).

Keywords

  • direct capture
  • polyclonal & monoclonal antibodies
  • 4-mercaptoethylpyridine
  • serum & plasma
  • complex unclarified bioprocess liquors

Cite this

@article{01c3411213ae4953b4d3d6e2e48aef5d,
title = "Magnetic hydrophobic-charge induction adsorbents for the recovery of immunoglobulins from antiserum feedstocks by high-gradient magnetic fishing",
abstract = "BACKGROUND The extraction of biopharmaceuticals from plasma and serum often employs overly complicated antiquated procedures, that can inflict serious damage on especially prone protein targets and which afford low purification power and overall yields. Here we describe systematic development of a high-gradient magnetic fishing process for recovery of immunoglobulins from unclarified antiserum. RESULTS Non-porous superparamagnetic particles were transformed into hydrophobic-charge induction adsorbents and then used to recover immunoglobulins from rabbit antiserum feedstocks. Comprehensive characterisation tests conducted with variously diluted clarified antiserum on a magnetic rack revealed that immunoglobulin binding was rapid (equilibrium reached in <45 s), strong (Kd <0.1 mg mL-1), of high capacity (Qmax = 214 mg g-1), and pH and ionic strength dependent. In a high-gradient magnetic fishing process conducted with the same adsorbent, and a conventional ‘magnetic filter + recycle loop’ arrangement, >72{\%} of the immunoglobulin present in an unclarified antiserum feed was recovered in 0.5 h in >3-fold purified form. CONCLUSIONS Fast magnetic particle based capture of antibodies from an unclarified high-titre feed has been demonstrated. Efficient product recovery from ultra-high titre bioprocess liquors by high-gradient magnetic fishing requires that improved magnetic adsorbents displaying high selectivity, ultra-high capacity and operational robustness are used with 'state-of-the-art’ rotor-stator magnetic separators.",
keywords = "direct capture, polyclonal & monoclonal antibodies, 4-mercaptoethylpyridine, serum & plasma, complex unclarified bioprocess liquors",
author = "Gomes, {Cl{\'a}udia Sg} and Adedayo Fashina and Alfred Fern{\'a}ndez-castan{\'e} and Overton, {Timothy W} and Hobley, {Timothy J} and Eirini Theodosiou and Thomas, {Owen Rt}",
note = "Funding: Biotechnology & Biological Sciences Research Council (BBSRC) and the Danish Council forS trategic Research( DetStrategiskeForskningsr{\aa}d).",
year = "2018",
month = "2",
day = "2",
doi = "10.1002/jctb.5599",
language = "English",
journal = "Journal of Chemical Technology and Biotechnology",
issn = "0268-2575",
publisher = "John Wiley and Sons Ltd",

}

Magnetic hydrophobic-charge induction adsorbents for the recovery of immunoglobulins from antiserum feedstocks by high-gradient magnetic fishing. / Gomes, Cláudia Sg; Fashina, Adedayo; Fernández-castané, Alfred; Overton, Timothy W; Hobley, Timothy J; Theodosiou, Eirini; Thomas, Owen Rt.

In: Journal of Chemical Technology and Biotechnology, 02.02.2018.

Research output: Contribution to journalArticle

TY - JOUR

T1 - Magnetic hydrophobic-charge induction adsorbents for the recovery of immunoglobulins from antiserum feedstocks by high-gradient magnetic fishing

AU - Gomes, Cláudia Sg

AU - Fashina, Adedayo

AU - Fernández-castané, Alfred

AU - Overton, Timothy W

AU - Hobley, Timothy J

AU - Theodosiou, Eirini

AU - Thomas, Owen Rt

N1 - Funding: Biotechnology & Biological Sciences Research Council (BBSRC) and the Danish Council forS trategic Research( DetStrategiskeForskningsråd).

PY - 2018/2/2

Y1 - 2018/2/2

N2 - BACKGROUND The extraction of biopharmaceuticals from plasma and serum often employs overly complicated antiquated procedures, that can inflict serious damage on especially prone protein targets and which afford low purification power and overall yields. Here we describe systematic development of a high-gradient magnetic fishing process for recovery of immunoglobulins from unclarified antiserum. RESULTS Non-porous superparamagnetic particles were transformed into hydrophobic-charge induction adsorbents and then used to recover immunoglobulins from rabbit antiserum feedstocks. Comprehensive characterisation tests conducted with variously diluted clarified antiserum on a magnetic rack revealed that immunoglobulin binding was rapid (equilibrium reached in <45 s), strong (Kd <0.1 mg mL-1), of high capacity (Qmax = 214 mg g-1), and pH and ionic strength dependent. In a high-gradient magnetic fishing process conducted with the same adsorbent, and a conventional ‘magnetic filter + recycle loop’ arrangement, >72% of the immunoglobulin present in an unclarified antiserum feed was recovered in 0.5 h in >3-fold purified form. CONCLUSIONS Fast magnetic particle based capture of antibodies from an unclarified high-titre feed has been demonstrated. Efficient product recovery from ultra-high titre bioprocess liquors by high-gradient magnetic fishing requires that improved magnetic adsorbents displaying high selectivity, ultra-high capacity and operational robustness are used with 'state-of-the-art’ rotor-stator magnetic separators.

AB - BACKGROUND The extraction of biopharmaceuticals from plasma and serum often employs overly complicated antiquated procedures, that can inflict serious damage on especially prone protein targets and which afford low purification power and overall yields. Here we describe systematic development of a high-gradient magnetic fishing process for recovery of immunoglobulins from unclarified antiserum. RESULTS Non-porous superparamagnetic particles were transformed into hydrophobic-charge induction adsorbents and then used to recover immunoglobulins from rabbit antiserum feedstocks. Comprehensive characterisation tests conducted with variously diluted clarified antiserum on a magnetic rack revealed that immunoglobulin binding was rapid (equilibrium reached in <45 s), strong (Kd <0.1 mg mL-1), of high capacity (Qmax = 214 mg g-1), and pH and ionic strength dependent. In a high-gradient magnetic fishing process conducted with the same adsorbent, and a conventional ‘magnetic filter + recycle loop’ arrangement, >72% of the immunoglobulin present in an unclarified antiserum feed was recovered in 0.5 h in >3-fold purified form. CONCLUSIONS Fast magnetic particle based capture of antibodies from an unclarified high-titre feed has been demonstrated. Efficient product recovery from ultra-high titre bioprocess liquors by high-gradient magnetic fishing requires that improved magnetic adsorbents displaying high selectivity, ultra-high capacity and operational robustness are used with 'state-of-the-art’ rotor-stator magnetic separators.

KW - direct capture

KW - polyclonal & monoclonal antibodies

KW - 4-mercaptoethylpyridine

KW - serum & plasma

KW - complex unclarified bioprocess liquors

U2 - 10.1002/jctb.5599

DO - 10.1002/jctb.5599

M3 - Article

JO - Journal of Chemical Technology and Biotechnology

JF - Journal of Chemical Technology and Biotechnology

SN - 0268-2575

ER -