Camelid VHH affinity ligands enable separation of closely related biopharmaceuticals.
Interest in new and diverse classes of molecules such as recombinant toxins, enzymes, and blood factors continues to grow for use a biotherapeutics. Compared to monoclonal antibodies, these novel drugs typically lack a commercially available affinity chromatography option, which leads to greater process complexity, longer development timelines, and poor platformability. To date, for both monoclonal antibodies and novel molecules, affinity chromatography has been mostly reserved for separation of process-related impurities such as host cell proteins and DNA. Reports of affinity purification of closely related product variants and modified forms are much rarer. In this work we describe custom affinity chromatography development using camelid VHH antibody fragments as "tunable" immunoaffinity ligands for separation of product-related impurities. One example demonstrates high selectivity for a recombinant immunotoxin where no binding was observed for an undesired deamidated species. Also discussed is affinity purification of a coagulation factor through specific recognition of the gamma-carboxylglutamic acid domain.
MedImmune, Department of Purification Process Sciences, Gaithersburg, MD, USA.
Timothy M
TM
Pabst
MedImmune, Department of Purification Process Sciences, Gaithersburg, MD, USA.
Michaela
M
Wendeler
MedImmune, Department of Purification Process Sciences, Gaithersburg, MD, USA.
Xiangyang
X
Wang
Thermo Fisher Scientific, Naarden, The Netherlands.
Sandra
S
Bezemer
Thermo Fisher Scientific, Naarden, The Netherlands.
Pim
P
Hermans
MedImmune, Department of Purification Process Sciences, Gaithersburg, MD, USA.
Alan K
AK
Hunter
D000906:Antibodies / Q000302:isolation & purification; D001427:Bacterial Toxins / Q000302:isolation & purification; D002846:Chromatography, Affinity / Q000379:methods*; D005098:Exotoxins / Q000302:isolation & purification; D011516:Prothrombin / Q000302:isolation & purification
D000906:Antibodies; D001427:Bacterial Toxins; D005098:Exotoxins; C541974:immunotoxin HA22; D011516:Prothrombin
Affinity chromatography; Conformational epitopes; Moxetumomab pasudotox; Prothrombin; VHH ligand
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