Spider silk fibers spun from soluble recombinant silk produced in mammalian cells.
Spider silks are protein-based "biopolymer" filaments or threads secreted by specialized epithelial cells as concentrated soluble precursors of highly repetitive primary sequences. Spider dragline silk is a flexible, lightweight fiber of extraordinary strength and toughness comparable to that of synthetic high-performance fibers. We sought to "biomimic" the process of spider silk production by expressing in mammalian cells the dragline silk genes (ADF-3/MaSpII and MaSpI) of two spider species. We produced soluble recombinant (rc)-dragline silk proteins with molecular masses of 60 to 140 kilodaltons. We demonstrated the wet spinning of silk monofilaments spun from a concentrated aqueous solution of soluble rc-spider silk protein (ADF-3; 60 kilodaltons) under modest shear and coagulation conditions. The spun fibers were water insoluble with a fine diameter (10 to 40 micrometers) and exhibited toughness and modulus values comparable to those of native dragline silks but with lower tenacity. Dope solutions with rc-silk protein concentrations >20% and postspinning draw were necessary to achieve improved mechanical properties of the spun fibers. Fiber properties correlated with finer fiber diameter and increased birefringence.
Lazaris
Anthoula
A
Nexia Biotechnologies, Vaudreuil-Dorion, Quebec J7V 8P5, Canada. alazaris@nexiabiotech.com
Arcidiacono
Steven
S
Huang
Yue
Y
Zhou
Jiang-Feng
JF
Duguay
Francois
F
Chretien
Nathalie
N
Welsh
Elizabeth A
EA
Soares
Jason W
JW
Karatzas
Costas N
CN
D020816:Amino Acid Motifs; D000595:Amino Acid Sequence; D000818:Animals; D001704:Biopolymers; D001718:Birefringence; D002417:Cattle; D002460:Cell Line; D003001:Cloning, Molecular; D006224:Cricetinae; D017077:Culture Media, Conditioned; D018076:DNA, Complementary; D004548:Elasticity; D004847:Epithelial Cells / Q000378:metabolism; D005349:Fibroins; D008422:Materials Testing; D019563:Mechanics; D008969:Molecular Sequence Data; D008970:Molecular Weight; D014176:Protein Biosynthesis; D017433:Protein Structure, Secondary; D011506:Proteins / Q000737:chemistry / Q000235:genetics* / Q000302:isolation & purification; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000737:chemistry / Q000302:isolation & purification; D012995:Solubility; D013112:Spiders / Q000235:genetics* / Q000378:metabolism; D013314:Stress, Mechanical; D013718:Tensile Strength; D014867:Water
D001704:Biopolymers; D017077:Culture Media, Conditioned; D018076:DNA, Complementary; D011506:Proteins; D011994:Recombinant Proteins; D014867:Water; C076815:spidroin 2; D005349:Fibroins