title: Evaluation of strategies to control Fab light chain dimer during mammalian expression and purification: A universal one-step process for purification of correctly assembled Fab.
abstract: Fabs are an important class of antibody fragment as both research reagents and therapeutic agents. There are a plethora of methods described for their recombinant expression and purification. However, these do not address the issue of excessive light chain production that forms light chain dimers nor do they describe a universal purification strategy. Light chain dimer impurities and the absence of a universal Fab purification strategy present persistent challenges for biotechnology applications using Fabs, particularly around the need for bespoke purification strategies. This study describes methods to address light chain dimer formation during Fab expression and identifies a novel CH 1 affinity resin as a simple and efficient one-step purification for correctly assembled Fab.
authors.affiliation: MedImmune, Granta Park, Cambridge, United Kingdom.
authors.forename: Jennifer
authors.initials: J
authors.lastname: Spooner
authors.affiliation: MedImmune, Granta Park, Cambridge, United Kingdom.
authors.forename: Jenny
authors.initials: J
authors.lastname: Keen
authors.affiliation: MedImmune, Granta Park, Cambridge, United Kingdom.
authors.forename: Kalpana
authors.initials: K
authors.lastname: Nayyar
authors.affiliation: MedImmune, Granta Park, Cambridge, United Kingdom.
authors.forename: Neil
authors.initials: N
authors.lastname: Birkett
authors.affiliation: MedImmune, Granta Park, Cambridge, United Kingdom.
authors.forename: Nicholas
authors.initials: N
authors.lastname: Bond
authors.affiliation: MedImmune, Granta Park, Cambridge, United Kingdom.
authors.forename: David
authors.initials: D
authors.lastname: Bannister
authors.affiliation: MedImmune, Granta Park, Cambridge, United Kingdom.
authors.forename: Natalie
authors.initials: N
authors.lastname: Tigue
authors.affiliation: MedImmune, Granta Park, Cambridge, United Kingdom.
authors.forename: Daniel
authors.initials: D
authors.lastname: Higazi
authors.affiliation: MedImmune, Granta Park, Cambridge, United Kingdom.
authors.forename: Benjamin
authors.initials: B
authors.lastname: Kemp
authors.affiliation: MedImmune, Granta Park, Cambridge, United Kingdom.
authors.forename: Tristan
authors.initials: T
authors.lastname: Vaughan
authors.affiliation: MedImmune, Granta Park, Cambridge, United Kingdom.
authors.forename: Alistair
authors.initials: A
authors.lastname: Kippen
authors.affiliation: MedImmune, Granta Park, Cambridge, United Kingdom. buchanana@medimmune.com.
authors.forename: Andrew
authors.initials: A
authors.lastname: Buchanan
mesh_terms: D019281:Dimerization; D015870:Gene Expression; D007140:Immunoglobulin Fab Fragments / Q000096:biosynthesis* / Q000235:genetics / Q000302:isolation & purification*; D007147:Immunoglobulin Light Chains / Q000096:biosynthesis* / Q000235:genetics / Q000302:isolation & purification*; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000235:genetics
chemical_list: D007140:Immunoglobulin Fab Fragments; D007147:Immunoglobulin Light Chains; D011994:Recombinant Proteins
keywords: antibody fragment; expression; fab; light chain dimer; purification