DNA aptamer affinity ligands for highly selective purification of human plasma-related proteins from multiple sources.
Nucleic acid aptamers are promising ligands for analytical and preparative-scale affinity chromatography applications. However, a full industrial exploitation requires that aptamer-grafted chromatography media provide a number of high technical standards that remained largely untested. Ideally, they should exhibit relatively high binding capacity associated to a very high degree of specificity. In addition, they must be highly resistant to harsh cleaning/sanitization conditions, as well as to prolonged and repeated exposure to biological environment. Here, we present practical examples of aptamer affinity chromatography for the purification of three human therapeutic proteins from various sources: Factor VII, Factor H and Factor IX. In a single chromatographic step, three DNA aptamer ligands enabled the efficient purification of their target protein, with an unprecedented degree of selectivity (from 0.5% to 98% of purity in one step). Furthermore, these aptamers demonstrated a high stability under harsh sanitization conditions (100h soaking in 1M NaOH). These results pave the way toward a wider adoption of aptamer-based affinity ligands in the industrial-scale purification of not only plasma-derived proteins but also of any other protein in general.
LFB Biotechnologies, Les Ulis, France.
Cynthia
C
Forier
Bioconsultant, JAM Conseil, Neuilly, France.
Egisto
E
Boschetti
INSERM U1023, LIME, Orsay, France.
Mohamed
M
Ouhammouch
INSERM U1023, LIME, Orsay, France.
Agnès
A
Cibiel
CEA, I2BM, MIRCen, UMR 9199, Université Paris Saclay, Fontenay aux Roses, France.
Frédéric
F
Ducongé
LFB Biotechnologies, Les Ulis, France.
Michel
M
Nogré
LFB Biotechnologies, Les Ulis, France.
Michel
M
Tellier
LFB Biotechnologies, Les Ulis, France.
Damien
D
Bataille
LFB Biotechnologies, Les Ulis, France.
Nicolas
N
Bihoreau
LFB Biotechnologies, Les Ulis, France.
Patrick
P
Santambien
LFB Biotechnologies, Les Ulis, France.
Sami
S
Chtourou
LFB Biotechnologies, Les Ulis, France. Electronic address: perretg@lfb.fr.
Gérald
G
Perret
D052157:Aptamers, Nucleotide / Q000737:chemistry; D001798:Blood Proteins / Q000302:isolation & purification*; D002846:Chromatography, Affinity / Q000379:methods*; D004247:DNA / Q000737:chemistry; D006801:Humans; D008024:Ligands
D052157:Aptamers, Nucleotide; D001798:Blood Proteins; D008024:Ligands; D004247:DNA
Affinity ligands; Aptamers; Blood proteins; Chromatography; Selectivity