Fibroin peptide and protein fragment compositions

Abstract

The present disclosure relates to peptide compositions, for example, silk fibroin-derived peptide compositions.

Description

[Technical Field]

[0001] The present disclosure relates to peptide compositions, such as silk fibroin-derived peptide compositions. The present disclosure is also in the field of synthetic textiles coated with silk fibroin proteins and protein fragments. In further embodiments, the present disclosure is in the field of silk fibroin compositions and methods for stimulating collagen and / or claudin-1 expression. [Background technology]

[0002] Silk is a natural polymer produced by a variety of insects and spiders and comprises a filament core protein, silk fibroin, and a colloidal coating consisting of the non-filamentous protein, sericin. Summary of the Invention

[0003] The present disclosure provides peptide compositions, eg, silk fibroin-derived peptide compositions.

[0004] The present disclosure provides peptides or protein fragments comprising a plurality of amino acids selected from M, R, V, K, T, F, I, L, C, A, Q, Y, N, D, E, G, S, H, P, and W, wherein at least one of the amino acids is modified, substituted, or replaced. In some embodiments, the peptide or protein fragment is a peptide or protein fragment of fibroin comprising an amino acid modification, substitution, or replacement of an amino acid from an amino acid selected from M, R, V, K, T, F, I, L, C, A, Q, Y, N, D, E, G, S, H, P, and W. In some embodiments, the fibroin is a fibroin heavy chain, a fibroin light chain, or a fibrohexamerin. In some embodiments, the peptide or protein fragment comprises from about 2 to about 100 amino acids. In some embodiments, the peptide or protein fragment comprises from about 2 to about 25 amino acids. In some embodiments, the peptide or protein fragment comprises from about 25 to about 50 amino acids. In some embodiments, the peptide or protein fragment contains between about 50 and about 75 amino acids. In some embodiments, the peptide or protein fragment contains between about 75 and about 100 amino acids. In some embodiments, the peptide or protein fragment contains between about 100 and about 125 amino acids. In some embodiments, the peptide or protein fragment contains between about 125 and about 150 amino acids. In some embodiments, the peptide or protein fragment contains between about 150 and about 200 amino acids. In some embodiments, the peptide or protein fragment contains between about 200 and about 250, 300, 350, 400, 450, or 500 amino acids. In some embodiments, the peptide or protein fragment contains between 1 and 5 modifications, substitutions, and / or substitutions. In some embodiments, the peptide or protein fragment contains one modification, substitution, and / or substitution. In some embodiments, the peptide or protein fragment contains two modifications, substitutions, and / or substitutions. In some embodiments, the peptide or protein fragment contains three modifications, substitutions, and / or substitutions.In some embodiments, the peptide or protein fragment comprises four modifications, substitutions, and / or substitutions. In some embodiments, the peptide or protein fragment comprises five modifications, substitutions, and / or substitutions. In some embodiments, the peptide or protein fragment comprises six modifications, substitutions, and / or substitutions. In some embodiments, the peptide or protein fragment comprises seven modifications, substitutions, and / or substitutions. In some embodiments, the peptide or protein fragment comprises eight modifications, substitutions, and / or substitutions. In some embodiments, the peptide or protein fragment comprises nine modifications, substitutions, and / or substitutions. In some embodiments, the peptide or protein fragment comprises ten modifications, substitutions, and / or substitutions. In some embodiments, the modifications, substitutions, and / or substitutions are selected from an asparagine to aspartic acid modification, substitution, and / or substitution, a glutamine to glutamic acid modification, substitution, and / or substitution, and a methionine to methionine oxide modification, substitution, and / or substitution. In some embodiments, the fibroin is a fibroin heavy chain and the modification, substitution, and / or substitution is at a position corresponding to any one of positions 1-5263 of the fibroin heavy chain. In some embodiments, the modification, substitution, and / or substitution is at positions Q58, M64, N68, N70, N77, M80, N93, M103, Q125, N132, Q139, Q275, N4191, Q5216, and / or N5262 of the fibroin heavy chain. In some embodiments, the fibroin is a fibroin light chain and the modification, substitution, and / or substitution is at a position corresponding to any one of positions 1-262 of the fibroin light chain. In some embodiments, the modifications, substitutions, and / or substitutions are at positions N23, Q24, N28, M69, N105, N108, N118, N136, N138, Q149, N186, N200, Q202, N204, N240, N248, and / or Q255 of the fibroin light chain.In some embodiments, the fibroin is fibrohexamerin (p25) and the modification, substitution, and / or substitution is at a position corresponding to any one of positions 1 to 220 of fibrohexamerin (p25). In some embodiments, the modification, substitution, and / or substitution is at positions Q62, N93, M120, N149, N172, N174, and / or N202 of fibrohexamerin (p25).

[0005] The present disclosure provides compositions comprising a plurality of peptides or protein fragments, each peptide or protein fragment comprising a plurality of amino acids selected from M, R, V, K, T, F, I, L, C, A, Q, Y, N, D, E, G, S, H, P, and W, wherein at least one of the amino acids is modified, substituted, or replaced. In some embodiments, the plurality of peptides or protein fragments comprises peptides or protein fragments of fibroin comprising amino acid modifications, substitutions, or replacements of amino acids selected from M, R, V, K, T, F, I, L, C, A, Q, Y, N, D, E, G, S, H, P, and W. In some embodiments, the fibroin is a fibroin heavy chain, a fibroin light chain, or a fibrohexamerin. In some embodiments, the peptide or protein fragment comprises from about 2 to about 100 amino acids. In some embodiments, the peptide or protein fragment comprises from about 2 to about 25 amino acids. In some embodiments, the peptide or protein fragment comprises from about 25 to about 50 amino acids. In some embodiments, the peptide or protein fragment contains between about 50 and about 75 amino acids. In some embodiments, the peptide or protein fragment contains between about 75 and about 100 amino acids. In some embodiments, the peptide or protein fragment contains between about 100 and about 125 amino acids. In some embodiments, the peptide or protein fragment contains between about 125 and about 150 amino acids. In some embodiments, the peptide or protein fragment contains between about 150 and about 200 amino acids. In some embodiments, the peptide or protein fragment contains between about 200 and about 250, 300, 350, 400, 450, or 500 amino acids. In some embodiments, the peptide or protein fragment contains between 1 and 5 modifications, substitutions, and / or substitutions. In some embodiments, the peptide or protein fragment contains one modification, substitution, and / or substitution. In some embodiments, the peptide or protein fragment contains two modifications, substitutions, and / or substitutions. In some embodiments, the peptide or protein fragment contains three modifications, substitutions, and / or substitutions.In some embodiments, the peptide or protein fragment comprises four modifications, substitutions, and / or substitutions. In some embodiments, the peptide or protein fragment comprises five modifications, substitutions, and / or substitutions. In some embodiments, the peptide or protein fragment comprises six modifications, substitutions, and / or substitutions. In some embodiments, the peptide or protein fragment comprises seven modifications, substitutions, and / or substitutions. In some embodiments, the peptide or protein fragment comprises eight modifications, substitutions, and / or substitutions. In some embodiments, the peptide or protein fragment comprises nine modifications, substitutions, and / or substitutions. In some embodiments, the peptide or protein fragment comprises ten modifications, substitutions, and / or substitutions. In some embodiments, the modifications, substitutions, and / or substitutions are selected from an asparagine to aspartic acid modification, substitution, and / or substitution, a glutamine to glutamic acid modification, substitution, and / or substitution, and a methionine to methionine oxide modification, substitution, and / or substitution. In some embodiments, the fibroin is a fibroin heavy chain and the modification, substitution, and / or substitution is at a position corresponding to any one of positions 1-5263 of the fibroin heavy chain. In some embodiments, the modification, substitution, and / or substitution is at positions Q58, M64, N68, N70, N77, M80, N93, M103, Q125, N132, Q139, Q275, N4191, Q5216, and / or N5262 of the fibroin heavy chain. In some embodiments, the fibroin is a fibroin light chain and the modification, substitution, and / or substitution is at a position corresponding to any one of positions 1-262 of the fibroin light chain. In some embodiments, the modifications, substitutions, and / or substitutions are at positions N23, Q24, N28, M69, N105, N108, N118, N136, N138, Q149, N186, N200, Q202, N204, N240, N248, and / or Q255 of the fibroin light chain.In some embodiments, the fibroin is fibrohexamerin (p25), and the modification, substitution, and / or substitution is at a position corresponding to any one of positions 1-220 of fibrohexamerin (p25). In some embodiments, the modification, substitution, and / or substitution is at positions Q62, N93, M120, N149, N172, N174, and / or N202 of fibrohexamerin (p25). In some embodiments, each modification, substitution, and / or substitution independently accounts for about 1% to about 99% of the composition. In some embodiments, each modification, substitution, and / or substitution independently accounts for about 1% to about 10% of the composition. In some embodiments, each modification, substitution, and / or substitution independently accounts for about 10% to about 20% of the composition. In some embodiments, each modification, substitution, and / or substitution independently accounts for about 20% to about 30% of the composition. In some embodiments, each modification, substitution, and / or substitution independently accounts for about 30% to about 40% of the composition. In some embodiments, each modification, substitution, and / or substitution independently accounts for about 40% to about 50% of the composition. In some embodiments, each modification, substitution, and / or substitution independently accounts for about 50% to about 60% of the composition. In some embodiments, each modification, substitution, and / or substitution independently accounts for about 60% to about 70% of the composition. In some embodiments, each modification, substitution, and / or substitution independently accounts for about 70% to about 80% of the composition. In some embodiments, each modification, substitution, and / or substitution independently accounts for about 80% to about 90% of the composition. In some embodiments, each modification, substitution, and / or substitution independently accounts for about 90% to about 99% of the composition, or about 90% to about 99% of the composition. As used herein, % modification, substitution, and / or substitution is defined as (the number of peptides or protein fragments containing a modification, substitution, and / or substitution at a particular position divided by the total number of peptides or protein fragments containing the particular position, whether or not they contain the modification, substitution, and / or substitution) x 100.

[0006] The present disclosure provides compositions comprising a plurality of peptide or protein fragments of fibroin heavy chain, fibroin light chain, and / or fibrohexamerin (p25), the compositions comprising one or more fractions, wherein the plurality of peptide or protein fragments comprises peptide or protein fragments of fibroin that include amino acid modifications, substitutions, or replacements. In some embodiments, the plurality of peptides or protein fragments have a weight average molecular weight (M) selected from about 1 kDa to about 20 kDa, about 20 kDa to about 40 kDa, about 40 kDa to about 60 kDa, about 60 kDa to about 80 kDa, about 80 kDa to about 100 kDa, about 100 kDa to about 120 kDa, about 120 kDa to about 140 kDa, or about 140 kDa to about 160 kDa, about 160 kDa to about 180 kDa, about 180 kDa to about 200 kDa, or about 200 kDa to about 250 kDa. w ), and a polydispersity of 1 to about 1.7, 1 to less than 3, or 1 to less than 5. In some embodiments, the plurality of peptides or protein fragments in the fraction have a weight average molecular weight (M) selected from about 10 kDa to about 20 kDa, about 20 kDa to about 40 kDa, about 40 kDa to about 60 kDa, about 60 kDa to about 80 kDa, about 80 kDa to about 100 kDa, about 100 kDa to about 120 kDa, about 120 kDa to about 140 kDa, about 140 kDa to about 160 kDa, or about 160 kDa to about 180 kDa. w ), and a polydispersity of 1 to about 1.1, or 1 to about 1.2, or 1 to less than 3, or 1 to less than 5. In some embodiments, the plurality of peptides or protein fragments in the fraction have a weight average molecular weight (Mw) selected from about 10 kDa to about 20 kDa, about 20 kDa to about 40 kDa, about 40 kDa to about 60 kDa, about 60 kDa to about 80 kDa, about 80 kDa to about 100 kDa, about 100 kDa to about 120 kDa, or about 120 kDa to about 140 kDa, and a polydispersity of 1 to about 1.1, or 1 to about 1.2, or 1 to less than 3, or 1 to less than 5. In some embodiments, the plurality of peptides or protein fragments in the fraction have a weight average molecular weight (M) selected from about 60 kDa to about 80 kDa, about 80 kDa to about 100 kDa, or about 100 kDa to about 120 kDa. w), and a polydispersity of 1 to about 1.1, or 1 to 3, or 1 to less than 5. In some embodiments, the plurality of peptides or protein fragments in the fraction have a weight average molecular weight (Mw) selected from about 10 kDa to about 20 kDa, about 20 kDa to about 40 kDa, about 40 kDa to about 60 kDa, about 60 kDa to about 80 kDa, about 80 kDa to about 100 kDa, or about 100 kDa to about 110 kDa, and a polydispersity of 1 to about 1.1, or 1 to about 1.2, or 1 to less than 3, or 1 to less than 5. In some embodiments, the plurality of peptides or protein fragments in the fraction have a weight average molecular weight (M) selected from about 60 kDa to about 80 kDa, about 80 kDa to about 100 kDa, about 100 kDa to about 120 kDa, or about 120 kDa to about 140 kDa. w ), and a polydispersity of 1 to about 1.1, or 1 to 3, or less than 1 to 5. In some embodiments, the plurality of peptides or protein fragments in the fraction have a weight average molecular weight (M) selected from about 20 kDa to about 40 kDa, or about 40 kDa to about 60 kDa. w), and a polydispersity of 1 to about 1.1, or 1 to about 1.2, or 1 to less than 3, or 1 to less than 5. In some embodiments, one or more fractions are selected from AS77, AS78, AS79, AS80, and AS81. In some embodiments, one or more fractions are selected from AS82, AS83, AS84, AS85, AS86, AS87, AS88, and AS89. In some embodiments, one or more fractions are selected from AS90, AS91, AS92, AS93, and AS94. In some embodiments, one or more fractions are selected from AS95, AS96, AS97, AS98, AS99, and AS100. In some embodiments, the plurality of peptides or protein fragments have a weight average molecular weight (Mw) selected from about 40 kDa to about 60 kDa, about 60 kDa to about 80 kDa, about 80 kDa to about 100 kDa, about 100 kDa to about 120 kDa, about 120 kDa to about 140 kDa, about 140 kDa to about 160 kDa, about 160 kDa to about 180 kDa, about 180 kDa to about 200 kDa, or about 200 kDa to about 220 kDa, and a polydispersity of 1 to about 1.7, or 1 to less than 3, or 1 to less than 5. In some embodiments, the plurality of peptides or protein fragments in the fraction have a weight average molecular weight (Mw) selected from about 40 kDa to about 60 kDa, about 60 kDa to about 80 kDa, about 80 kDa to about 100 kDa, about 100 kDa to about 120 kDa, about 120 kDa to about 140 kDa, about 140 kDa to about 160 kDa, about 160 kDa to about 180 kDa, about 180 kDa to about 200 kDa, or about 200 kDa to about 210 kDa, and a polydispersity of 1 to about 1.2, or 1 to about 1.3, or 1 to less than 3, or 1 to less than 5. In some embodiments, the plurality of peptides or protein fragments in the fraction have a weight average molecular weight (M) selected from about 40 kDa to about 60 kDa, about 60 kDa to about 80 kDa, about 80 kDa to about 100 kDa, or about 100 kDa to about 110 kDa. w), and a polydispersity of 1 to about 1.1, or 1 to about 1.2, or 1 to less than 3, or 1 to less than 5. In some embodiments, a plurality of peptides or protein fragments in the fraction are in the about 60 kDa to about 80 kDa fraction and have a weight average molecular weight (Mw) selected from about 80 kDa to about 100 kDa, about 100 kDa to about 120 kDa, about 120 kDa to about 140 kDa, about 140 kDa to about 160 kDa, about 160 kDa to about 180 kDa, about 180 kDa to about 200 kDa, or about 200 kDa to about 210 kDa, and a polydispersity of 1 to about 1.2, or 1 to about 1.3, or 1 to less than 3, or 1 to less than 5. In some embodiments, one or more fractions are selected from AS101, AS102, AS103, AS104, and AS105. In some embodiments, one or more fractions are selected from AS106, AS107, AS108, AS109, AS110, and AS111. In some embodiments, the amino acids are selected from M, R, V, K, T, F, I, L, C, A, Q, Y, N, D, E, G, S, H, P, and W. In some embodiments, the peptide or protein fragment comprises about 2 to about 100 amino acids. In some embodiments, the peptide or protein fragment comprises about 2 to about 25 amino acids. In some embodiments, the peptide or protein fragment comprises about 25 to about 50 amino acids. In some embodiments, the peptide or protein fragment comprises about 50 to about 75 amino acids. In some embodiments, the peptide or protein fragment comprises about 75 to about 100 amino acids. In some embodiments, the peptide or protein fragment comprises about 100 to about 125 amino acids. In some embodiments, the peptide or protein fragment comprises about 125 to about 150 amino acids. In some embodiments, the peptide or protein fragment comprises about 150 to about 200 amino acids. In some embodiments, the peptide or protein fragment comprises about 200 to about 250, 300, 350, 400, 450, or 500 amino acids. In some embodiments, the peptide or protein fragment comprises 1 to 5 modifications, substitutions, and / or substitutions.In some embodiments, the peptide or protein fragment contains one modification, substitution, and / or substitution. In some embodiments, the peptide or protein fragment contains two modifications, substitutions, and / or substitutions. In some embodiments, the peptide or protein fragment contains three modifications, substitutions, and / or substitutions. In some embodiments, the peptide or protein fragment contains four modifications, substitutions, and / or substitutions. In some embodiments, the peptide or protein fragment contains five modifications, substitutions, and / or substitutions. In some embodiments, the peptide or protein fragment contains six modifications, substitutions, and / or substitutions. In some embodiments, the peptide or protein fragment contains seven modifications, substitutions, and / or substitutions. In some embodiments, the peptide or protein fragment contains eight modifications, substitutions, and / or substitutions. In some embodiments, the peptide or protein fragment contains nine modifications, substitutions, and / or substitutions. In some embodiments, the peptide or protein fragment contains ten modifications, substitutions, and / or substitutions. In some embodiments, the fibroin is a fibroin heavy chain and the modification, substitution, and / or substitution is at a position corresponding to any one of positions 1-5263 of the fibroin heavy chain. In some embodiments, the fibroin is a fibroin light chain and the modification, substitution, and / or substitution is at a position corresponding to any one of positions 1-262 of the fibroin light chain. In some embodiments, the fibroin is a fibrohexamerin (p25) and the modification, substitution, and / or substitution is at a position corresponding to any one of positions 1-220 of the fibrohexamerin (p25). In some embodiments, the modification, substitution, and / or substitution is selected from an asparagine to aspartic acid modification, substitution, and / or substitution, a glutamine to glutamic acid modification, substitution, and / or substitution, and a methionine to methionine oxide modification, substitution, and / or substitution.In some embodiments, the modification, substitution, and / or substitution is at a fibroin heavy chain position selected from Q58, M64, N68, N70, N77, M80, N93, M103, Q125, N132, Q139, Q275, N4191, Q5216, and / or N5262. In some embodiments, the modification, substitution, and / or substitution is at a fibroin light chain position selected from N23, Q24, N28, M69, N105, N108, N118, N136, N138, Q149, N186, N200, Q202, N204, N240, N248, and / or Q255. In some embodiments, the modification, substitution, and / or substitution is at a fibrohexamerin (p25) position selected from Q62, N93, M120, N149, N172, N174, and / or N202. In some embodiments, each modification, substitution, and / or substitution is independently present in about 1% to about 99% of the composition. In some embodiments, each modification, substitution, and / or substitution is independently present in about 1% to about 10% of the composition. In some embodiments, each modification, substitution, and / or substitution is independently present in about 10% to about 20% of the composition. In some embodiments, each modification, substitution, and / or substitution is independently present in about 20% to about 30% of the composition. In some embodiments, each modification, substitution, and / or substitution is independently present in about 30% to about 40% of the composition. In some embodiments, each modification, substitution, and / or substitution is independently present in about 40% to about 50% of the composition. In some embodiments, each modification, substitution, and / or substitution independently accounts for about 50% to about 60% of the composition. In some embodiments, each modification, substitution, and / or substitution independently accounts for about 60% to about 70% of the composition. In some embodiments, each modification, substitution, and / or substitution independently accounts for about 70% to about 80% of the composition. In some embodiments, each modification, substitution, and / or substitution independently accounts for about 80% to about 90% of the composition. In some embodiments, each modification, substitution. , and / or substitutions independently range from about 90% to about 99%, or from about 90% to about 99%, of the composition. As used herein, % modification, substitution, and / or substitution is defined as (the number of peptides or protein fragments containing the modification, substitution, and / or substitution at a particular position divided by the total number of peptides or protein fragments containing the particular position, whether or not they contain the modification, substitution, and / or substitution) × 100. In some embodiments, the molecular weight is determined by MALS.

[0007] The present disclosure provides an article comprising one or more peptides or protein fragments disclosed herein and / or one or more compositions disclosed herein. The composition may comprise an unmodified fibroin peptide and a protein fragment. The article may be selected from, but is not limited to, a personal care article disclosed herein, a coated textile disclosed herein, and / or an article for treating textiles disclosed herein (e.g., a laundry pod). The present disclosure provides an article comprising one or more peptides or protein fragments disclosed herein and / or one or more compositions disclosed herein and a substrate or any other supporting or combining element disclosed herein.

[0008] In some embodiments, the present disclosure provides compositions as disclosed herein that include a plurality of peptides or protein fragments, e.g., a plurality of fibroin heavy chain peptides or fibroin heavy chain fragments, and that include a plurality of amino acid modifications, substitutions, and / or substitutions, and further include, at specific positions, a ratio of one or more modifications, substitutions, and / or substitutions selected from the following: a Q58 to M64 modification, substitution, and / or replacement ratio of about 25:1, about 20:1, about 15:1, about 10:1, about 9:1, about 8:1, about 7:1, about 6:1, about 5:1, about 4:1, about 3:1, about 2:1, about 1:1, about 1:2, about 1:3, about 1:4, about 1:5, about 1:6, about 1:7, about 1:8, about 1:9, about 1:10, about 1:15, about 1:20, or about 1:25; a Q58 to N68 modification, substitution, and / or replacement ratio of about 25:1, about 20:1, about 15:1, about 10:1, about 9:1, about 8:1, about 7:1, about 6:1, about 5:1, about 4:1, about 3:1, about 2:1, about 1:1, about 1:2, about 1:3, about 1:4, about 1:5, about 1:6, about 1:7, about 1:8, about 1:9, about 1:10, about 1:15, about 1:20, or about 1:25; a Q58 to N70 modification, substitution, and / or replacement ratio of about 25:1, about 20:1, about 15:1, about 10:1, about 9:1, about 8:1, about 7:1, about 6:1, about 5:1, about 4:1, about 3:1, about 2:1, about 1:1, about 1:2, about 1:3, about 1:4, about 1:5, about 1:6, about 1:7, about 1:8, about 1:9, about 1:10, about 1:15, about 1:20, or about 1:25; a Q58 to N77 modification, substitution, and / or replacement ratio of about 25:1, about 20:1, about 15:1, about 10:1, about 9:1, about 8:1, about 7:1, about 6:1, about 5:1, about 4:1, about 3:1, about 2:1, about 1:1, about 1:2, about 1:3, about 1:4, about 1:5, about 1:6, about 1:7, about 1:8, about 1:9, about 1:10, about 1:15, about 1:20, or about 1:25; a Q58 to M80 modification, substitution, and / or replacement ratio of about 25:1, about 20:1, about 15:1, about 10:1, about 9:1, about 8:1, about 7:1, about 6:1, about 5:1, about 4:1, about 3:1, about 2:1, about 1:1, about 1:2, about 1:3, about 1:4, about 1:5, about 1:6, about 1:7, about 1:8, about 1:9, about 1:10, about 1:15, about 1:20, or about 1:25; a Q58 to N93 modification, substitution, and / or replacement ratio of about 25:1, about 20:1, about 15:1, about 10:1, about 9:1, about 8:1, about 7:1, about 6:1, about 5:1, about 4:1, about 3:1, about 2:1, about 1:1, about 1:2, about 1:3, about 1:4, about 1:5, about 1:6, about 1:7, about 1:8, about 1:9, about 1:10, about 1:15, about 1:20, or about 1:25; a Q58 to M103 modification, substitution, and / or replacement ratio of about 25:1, about 20:1, about 15:1, about 10:1, about 9:1, about 8:1, about 7:1, about 6:1, about 5:1, about 4:1, about 3:1, about 2:1, about 1:1, about 1:2, about 1:3, about 1:4, about 1:5, about 1:6, about 1:7, about 1:8, about 1:9, about 1:10, about 1:15, about 1:20, or about 1:25; a Q58 to Q125 modification, substitution, and / or replacement ratio of about 25:1, about 20:1, about 15:1, about 10:1, about 9:1, about 8:1, about 7:1, about 6:1, about 5:1, about 4:1, about 3:1, about 2:1, about 1:1, about 1:2, about 1:3, about 1:4, about 1:5, about 1:6, about 1:7, about 1:8, about 1:9, about 1:10, about 1:15, about 1:20, or about 1:25; a Q58 to N132 modification, substitution, and / or replacement ratio of about 25:1, about 20:1, about 15:1, about 10:1, about 9:1, about 8:1, about 7:1, about 6:1, about 5:1, about 4:1, about 3:1, about 2:1, about 1:1, about 1:2, about 1:3, about 1:4, about 1:5, about 1:6, about 1:7, about 1:8, about 1:9, about 1:10, about 1:15, about 1:20, or about 1:25; a Q58 to Q139 modification, substitution, and / or replacement ratio of about 25:1, about 20:1, about 15:1, about 10:1, about 9:1, about 8:1, about 7:1, about 6:1, about 5:1, about 4:1, about 3:1, about 2:1, about 1:1, about 1:2, about 1:3, about 1:4, about 1:5, about 1:6, about 1:7, about 1:8, about 1:9, about 1:10, about 1:15, about 1:20, or about 1:25; a Q58 to Q275 modification, substitution, and / or replacement ratio of about 25:1, about 20:1, about 15:1, about 10:1, about 9:1, about 8:1, about 7:1, about 6:1, about 5:1, about 4:1, about 3:1, about 2:1, about 1:1, about 1:2, about 1:3, about 1:4, about 1:5, about 1:6, about 1:7, about 1:8, about 1:9, about 1:10, about 1:15, about 1:20, or about 1:25; a modification, substitution, and / or substitution ratio of Q58 to N4191 of about 25:1, about 20:1, about 15:1, about 10:1, about 9:1, about 8:1, about 7:1, about 6:1, about 5:1, about 4:1, about 3:1, about 2:1, about 1:1, about 1:2, about 1:3, about 1:4, about 1:5, about 1:6, about 1:7, about 1:8, about 1:9, about 1:10, about 1:15, about 1:20, or about 1:25; a modification, substitution, and / or substitution ratio of Q58 to Q5216 of about 25:1, about 20:1, about 15:1, about 10:1, about 9:1, about 8:1, about 7:1, about 6:1, about 5:1, about 4:1, about 3:1, about 2:1, about 1:1, about 1:2, about 1:3, about 1:4, about 1:5, about 1:6, about 1:7, about 1:8, about 1:9, about 1:10, about 1:15, about 1:20, or about 1:25; a modification, substitution, and / or substitution ratio of Q58 to N5262 of about 25:1, about 20:1, about 15:1, about 10:1, about 9:1, about 8:1, about 7:1, about 6:1, about 5:1, about 4:1, about 3:1, about 2:1, about 1:1, about 1:2, about 1:3, about 1:4, about 1:5, about 1:6, about 1:7, about 1:8, about 1:9, about 1:10, about 1:15, about 1:20, or about 1:25; an M64 to N68 modification, substitution, and / or replacement ratio of about 25:1, about 20:1, about 15:1, about 10:1, about 9:1, about 8:1, about 7:1, about 6:1, about 5:1, about 4:1, about 3:1, about 2:1, about 1:1, about 1:2, about 1:3, about 1:4, about 1:5, about 1:6, about 1:7, about 1:8, about 1:9, about 1:10, about 1:15, about 1:20, or about 1:25; an M64 to N70 modification, substitution, and / or replacement ratio of about 25:1, about 20:1, about 15:1, about 10:1, about 9:1, about 8:1, about 7:1, about 6:1, about 5:1, about 4:1, about 3:1, about 2:1, about 1:1, about 1:2, about 1:3, about 1:4, about 1:5, about 1:6, about 1:7, about 1:8, about 1:9, about 1:10, about 1:15, about 1:20, or about 1:25; an M64 to N77 modification, substitution, and / or replacement ratio of about 25:1, about 20:1, about 15:1, about 10:1, about 9:1, about 8:1, about 7:1, about 6:1, about 5:1, about 4:1, about 3:1, about 2:1, about 1:1, about 1:2, about 1:3, about 1:4, about 1:5, about 1:6, about 1:7, about 1:8, about 1:9, about 1:10, about 1:15, about 1:20, or about 1:25; - a ratio of M64 to M80 modification, substitution, and / or replacement of about 25:1, about 20:1, about 15:1, about 10:1, about 9:1, about 8:1, about 7:1, about 6:1, about 5:1, about 4:1, about 3:1, about 2:1, about 1:1, about 1:2, about 1:3, about 1:4, about 1:5, about 1:6, about 1:7, about 1:8, about 1:9, about 1:10, about 1:15, about 1:20, or about 1:25; an M64 to N93 modification, substitution, and / or replacement ratio of about 25:1, about 20:1, about 15:1, about 10:1, about 9:1, about 8:1, about 7:1, about 6:1, about 5:1, about 4:1, about 3:1, about 2:1, about 1:1, about 1:2, about 1:3, about 1:4, about 1:5, about 1:6, about 1:7, about 1:8, about 1:9, about 1:10, about 1:15, about 1:20, or about 1:25; - a ratio of M64 to M103 modification, substitution, and / or replacement of about 25:1, about 20:1, about 15:1, about 10:1, about 9:1, about 8:1, about 7:1, about 6:1, about 5:1, about 4:1, about 3:1, about 2:1, about 1:1, about 1:2, about 1:3, about 1:4, about 1:5, about 1:6, about 1:7, about 1:8, about 1:9, about 1:10, about 1:15, about 1:20, or about 1:25; - a ratio of M64 to Q125 modification, substitution, and / or replacement of about 25:1, about 20:1, about 15:1, about 10:1, about 9:1, about 8:1, about 7:1, about 6:1, about 5:1, about 4:1, about 3:1, about 2:1, about 1:1, about 1:2, about 1:3, about 1:4, about 1:5, about 1:6, about 1:7, about 1:8, about 1:9, about 1:10, about 1:15, about 1:20, or about 1:25; - an M64 to N132 modification, substitution, and / or replacement ratio of about 25:1, about 20:1, about 15:1, about 10:1, about 9:1, about 8:1, about 7:1, about 6:1, about 5:1, about 4:1, about 3:1, about 2:1, about 1:1, about 1:2, about 1:3, about 1:4, about 1:5, about 1:6, about 1:7, about 1:8, about 1:9, about 1:10, about 1:15, about 1:20, or about 1:25; - a ratio of M64 to Q139 modifications, substitutions, and / or substitutions of about 25:1, about 20:1, about 15:1, about 10:1, about 9:1, about 8:1, about 7:1, about 6:1, about 5:1, about 4:1, about 3:1, about 2:1, about 1:1, about 1:2, about 1:3, about 1:4, about 1:5, about 1:6, about 1:7, about 1:8, about 1:9, about 1:10, about 1:15, about 1:20, or about 1:25; - a ratio of M64 to Q275 modification, substitution, and / or replacement of about 25:1, about 20:1, about 15:1, about 10:1, about 9:1, about 8:1, about 7:1, about 6:1, about 5:1, about 4:1, about 3:1, about 2:1, about 1:1, about 1:2, about 1:3, about 1:4, about 1:5, about 1:6, about 1:7, about 1:8, about 1:9, about 1:10, about 1:15, about 1:20, or about 1:25; - a ratio of M64 to N4191 modification, substitution, and / or replacement of about 25:1, about 20:1, about 15:1, about 10:1, about 9:1, about 8:1, about 7:1, about 6:1, about 5:1, about 4:1, about 3:1, about 2:1, about 1:1, about 1:2, about 1:3, about 1:4, about 1:5, about 1:6, about 1:7, about 1:8, about 1:9, about 1:10, about 1:15, about 1:20, or about 1:25; - a ratio of M64 to Q5216 modifications, substitutions, and / or substitutions of about 25:1, about 20:1, about 15:1, about 10:1, about 9:1, about 8:1, about 7:1, about 6:1, about 5:1, about 4:1, about 3:1, about 2:1, about 1:1, about 1:2, about 1:3, about 1:4, about 1:5, about 1:6, about 1:7, about 1:8, about 1:9, about 1:10, about 1:15, about 1:20, or about 1:25; - a ratio of M64 to N5262 modifications, substitutions, and / or substitutions of about 25:1, about 20:1, about 15:1, about 10:1, about 9:1, about 8:1, about 7:1, about 6:1, about 5:1, about 4:1, about 3:1, about 2:1, about 1:1, about 1:2, about 1:3, about 1:4, about 1:5, about 1:6, about 1:7, about 1:8, about 1:9, about 1:10, about 1:15, about 1:20, or about 1:25; an N68 to N70 modification, substitution, and / or replacement ratio of about 25:1, about 20:1, about 15:1, about 10:1, about 9:1, about 8:1, about 7:1, about 6:1, about 5:1, about 4:1, about 3:1, about 2:1, about 1:1, about 1:2, about 1:3, about 1:4, about 1:5, about 1:6, about 1:7, about 1:8, about 1:9, about 1:10, about 1:15, about 1:20, or about 1:25; an N68 to N77 modification, substitution, and / or replacement ratio of about 25:1, about 20:1, about 15:1, about 10:1, about 9:1, about 8:1, about 7:1, about 6:1, about 5:1, about 4:1, about 3:1, about 2:1, about 1:1, about 1:2, about 1:3, about 1:4, about 1:5, about 1:6, about 1:7, about 1:8, about 1:9, about 1:10, about 1:15, about 1:20, or about 1:25; an N68 to M80 modification, substitution, and / or replacement ratio of about 25:1, about 20:1, about 15:1, about 10:1, about 9:1, about 8:1, about 7:1, about 6:1, about 5:1, about 4:1, about 3:1, about 2:1, about 1:1, about 1:2, about 1:3, about 1:4, about 1:5, about 1:6, about 1:7, about 1:8, about 1:9, about 1:10, about 1:15, about 1:20, or about 1:25; an N93 modification, substitution, and / or substitution ratio of N68 of about 25:1, about 20:1, about 15:1, about 10:1, about 9:1, about 8:1, about 7:1, about 6:1, about 5:1, about 4:1, about 3:1, about 2:1, about 1:1, about 1:2, about 1:3, about 1:4, about 1:5, about 1:6, about 1:7, about 1:8, about 1:9, about 1:10, about 1:15, about 1:20, or about 1:25; an N68 to M103 modification, substitution, and / or replacement ratio of about 25:1, about 20:1, about 15:1, about 10:1, about 9:1, about 8:1, about 7:1, about 6:1, about 5:1, about 4:1, about 3:1, about 2:1, about 1:1, about 1:2, about 1:3, about 1:4, about 1:5, about 1:6, about 1:7, about 1:8, about 1:9, about 1:10, about 1:15, about 1:20, or about 1:25; an N68 to Q125 modification, substitution, and / or replacement ratio of about 25:1, about 20:1, about 15:1, about 10:1, about 9:1, about 8:1, about 7:1, about 6:1, about 5:1, about 4:1, about 3:1, about 2:1, about 1:1, about 1:2, about 1:3, about 1:4, about 1:5, about 1:6, about 1:7, about 1:8, about 1:9, about 1:10, about 1:15, about 1:20, or about 1:25; an N68 to N132 modification, substitution, and / or replacement ratio of about 25:1, about 20:1, about 15:1, about 10:1, about 9:1, about 8:1, about 7:1, about 6:1, about 5:1, about 4:1, about 3:1, about 2:1, about 1:1, about 1:2, about 1:3, about 1:4, about 1:5, about 1:6, about 1:7, about 1:8, about 1:9, about 1:10, about 1:15, about 1:20, or about 1:25; - a N68 to Q139 modification, substitution, and / or replacement ratio of about 25:1, about 20:1, about 15:1, about 10:1, about 9:1, about 8:1, about 7:1, about 6:1, about 5:1, about 4:1, about 3:1, about 2:1, about 1:1, about 1:2, about 1:3, about 1:4, about 1:5, about 1:6, about 1:7, about 1:8, about 1:9, about 1:10, about 1:15, about 1:20, or about 1:25; an N68 to Q275 modification, substitution, and / or replacement ratio of about 25:1, about 20:1, about 15:1, about 10:1, about 9:1, about 8:1, about 7:1, about 6:1, about 5:1, about 4:1, about 3:1, about 2:1, about 1:1, about 1:2, about 1:3, about 1:4, about 1:5, about 1:6, about 1:7, about 1:8, about 1:9, about 1:10, about 1:15, about 1:20, or about 1:25; a modification, substitution, and / or replacement ratio of N68 to N4191 of about 25:1, about 20:1, about 15:1, about 10:1, about 9:1, about 8:1, about 7:1, about 6:1, about 5:1, about 4:1, about 3:1, about 2:1, about 1:1, about 1:2, about 1:3, about 1:4, about 1:5, about 1:6, about 1:7, about 1:8, about 1:9, about 1:10, about 1:15, about 1:20, or about 1:25; a N68 to Q5216 modification, substitution, and / or substitution ratio of about 25:1, about 20:1, about 15:1, about 10:1, about 9:1, about 8:1, about 7:1, about 6:1, about 5:1, about 4:1, about 3:1, about 2:1, about 1:1, about 1:2, about 1:3, about 1:4, about 1:5, about 1:6, about 1:7, about 1:8, about 1:9, about 1:10, about 1:15, about 1:20, or about 1:25; a modification, substitution, and / or replacement ratio of N68 to N5262 of about 25:1, about 20:1, about 15:1, about 10:1, about 9:1, about 8:1, about 7:1, about 6:1, about 5:1, about 4:1, about 3:1, about 2:1, about 1:1, about 1:2, about 1:3, about 1:4, about 1:5, about 1:6, about 1:7, about 1:8, about 1:9, about 1:10, about 1:15, about 1:20, or about 1:25; an N70 to N77 modification, substitution, and / or replacement ratio of about 25:1, about 20:1, about 15:1, about 10:1, about 9:1, about 8:1, about 7:1, about 6:1, about 5:1, about 4:1, about 3:1, about 2:1, about 1:1, about 1:2, about 1:3, about 1:4, about 1:5, about 1:6, about 1:7, about 1:8, about 1:9, about 1:10, about 1:15, about 1:20, or about 1:25; an N70 to M80 modification, substitution, and / or replacement ratio of about 25:1, about 20:1, about 15:1, about 10:1, about 9:1, about 8:1, about 7:1, about 6:1, about 5:1, about 4:1, about 3:1, about 2:1, about 1:1, about 1:2, about 1:3, about 1:4, about 1:5, about 1:6, about 1:7, about 1:8, about 1:9, about 1:10, about 1:15, about 1:20, or about 1:25; an N70 to N93 modification, substitution, and / or replacement ratio of about 25:1, about 20:1, about 15:1, about 10:1, about 9:1, about 8:1, about 7:1, about 6:1, about 5:1, about 4:1, about 3:1, about 2:1, about 1:1, about 1:2, about 1:3, about 1:4, about 1:5, about 1:6, about 1:7, about 1:8, about 1:9, about 1:10, about 1:15, about 1:20, or about 1:25; an N70 to M103 modification, substitution, and / or replacement ratio of about 25:1, about 20:1, about 15:1, about 10:1, about 9:1, about 8:1, about 7:1, about 6:1, about 5:1, about 4:1, about 3:1, about 2:1, about 1:1, about 1:2, about 1:3, about 1:4, about 1:5, about 1:6, about 1:7, about 1:8, about 1:9, about 1:10, about 1:15, about 1:20, or about 1:25; an N70 to Q125 modification, substitution, and / or replacement ratio of about 25:1, about 20:1, about 15:1, about 10:1, about 9:1, about 8:1, about 7:1, about 6:1, about 5:1, about 4:1, about 3:1, about 2:1, about 1:1, about 1:2, about 1:3, about 1:4, about 1:5, about 1:6, about 1:7, about 1:8, about 1:9, about 1:10, about 1:15, about 1:20, or about 1:25; an N70 to N132 modification, substitution, and / or replacement ratio of about 25:1, about 20:1, about 15:1, about 10:1, about 9:1, about 8:1, about 7:1, about 6:1, about 5:1, about 4:1, about 3:1, about 2:1, about 1:1, about 1:2, about 1:3, about 1:4, about 1:5, about 1:6, about 1:7, about 1:8, about 1:9, about 1:10, about 1:15, about 1:20, or about 1:25; - an N70 to Q139 modification, substitution, and / or replacement ratio of about 25:1, about 20:1, about 15:1, about 10:1, about 9:1, about 8:1, about 7:1, about 6:1, about 5:1, about 4:1, about 3:1, about 2:1, about 1:1, about 1:2, about 1:3, about 1:4, about 1:5, about 1:6, about 1:7, about 1:8, about 1:9, about 1:10, about 1:15, about 1:20, or about 1:25; an N70 to Q275 modification, substitution, and / or replacement ratio of about 25:1, about 20:1, about 15:1, about 10:1, about 9:1, about 8:1, about 7:1, about 6:1, about 5:1, about 4:1, about 3:1, about 2:1, about 1:1, about 1:2, about 1:3, about 1:4, about 1:5, about 1:6, about 1:7, about 1:8, about 1:9, about 1:10, about 1:15, about 1:20, or about 1:25; a modification, substitution, and / or substitution ratio of N70 to N4191 of about 25:1, about 20:1, about 15:1, about 10:1, about 9:1, about 8:1, about 7:1, about 6:1, about 5:1, about 4:1, about 3:1, about 2:1, about 1:1, about 1:2, about 1:3, about 1:4, about 1:5, about 1:6, about 1:7, about 1:8, about 1:9, about 1:10, about 1:15, about 1:20, or about 1:25; - a N70 to Q5216 modification, substitution, and / or substitution ratio of about 25:1, about 20:1, about 15:1, about 10:1, about 9:1, about 8:1, about 7:1, about 6:1, about 5:1, about 4:1, about 3:1, about 2:1, about 1:1, about 1:2, about 1:3, about 1:4, about 1:5, about 1:6, about 1:7, about 1:8, about 1:9, about 1:10, about 1:15, about 1:20, or about 1:25; a modification, substitution, and / or substitution ratio of N70 to N5262 of about 25:1, about 20:1, about 15:1, about 10:1, about 9:1, about 8:1, about 7:1, about 6:1, about 5:1, about 4:1, about 3:1, about 2:1, about 1:1, about 1:2, about 1:3, about 1:4, about 1:5, about 1:6, about 1:7, about 1:8, about 1:9, about 1:10, about 1:15, about 1:20, or about 1:25; an N77 to M80 modification, substitution, and / or replacement ratio of about 25:1, about 20:1, about 15:1, about 10:1, about 9:1, about 8:1, about 7:1, about 6:1, about 5:1, about 4:1, about 3:1, about 2:1, about 1:1, about 1:2, about 1:3, about 1:4, about 1:5, about 1:6, about 1:7, about 1:8, about 1:9, about 1:10, about 1:15, about 1:20, or about 1:25; an N77 to N93 modification, substitution, and / or replacement ratio of about 25:1, about 20:1, about 15:1, about 10:1, about 9:1, about 8:1, about 7:1, about 6:1, about 5:1, about 4:1, about 3:1, about 2:1, about 1:1, about 1:2, about 1:3, about 1:4, about 1:5, about 1:6, about 1:7, about 1:8, about 1:9, about 1:10, about 1:15, about 1:20, or about 1:25; an N77 to M103 modification, substitution, and / or replacement ratio of about 25:1, about 20:1, about 15:1, about 10:1, about 9:1, about 8:1, about 7:1, about 6:1, about 5:1, about 4:1, about 3:1, about 2:1, about 1:1, about 1:2, about 1:3, about 1:4, about 1:5, about 1:6, about 1:7, about 1:8, about 1:9, about 1:10, about 1:15, about 1:20, or about 1:25; - a N77 to Q125 modification, substitution, and / or replacement ratio of about 25:1, about 20:1, about 15:1, about 10:1, about 9:1, about 8:1, about 7:1, about 6:1, about 5:1, about 4:1, about 3:1, about 2:1, about 1:1, about 1:2, about 1:3, about 1:4, about 1:5, about 1:6, about 1:7, about 1:8, about 1:9, about 1:10, about 1:15, about 1:20, or about 1:25; - a N77 to N132 modification, substitution, and / or replacement ratio of about 25:1, about 20:1, about 15:1, about 10:1, about 9:1, about 8:1, about 7:1, about 6:1, about 5:1, about 4:1, about 3:1, about 2:1, about 1:1, about 1:2, about 1:3, about 1:4, about 1:5, about 1:6, about 1:7, about 1:8, about 1:9, about 1:10, about 1:15, about 1:20, or about 1:25; a N77 to Q139 modification, substitution, and / or replacement ratio of about 25:1, about 20:1, about 15:1, about 10:1, about 9:1, about 8:1, about 7:1, about 6:1, about 5:1, about 4:1, about 3:1, about 2:1, about 1:1, about 1:2, about 1:3, about 1:4, about 1:5, about 1:6, about 1:7, about 1:8, about 1:9, about 1:10, about 1:15, about 1:20, or about 1:25; a N77 to Q275 modification, substitution, and / or replacement ratio of about 25:1, about 20:1, about 15:1, about 10:1, about 9:1, about 8:1, about 7:1, about 6:1, about 5:1, about 4:1, about 3:1, about 2:1, about 1:1, about 1:2, about 1:3, about 1:4, about 1:5, about 1:6, about 1:7, about 1:8, about 1:9, about 1:10, about 1:15, about 1:20, or about 1:25; a modification, substitution, and / or replacement ratio of N77 to N4191 of about 25:1, about 20:1, about 15:1, about 10:1, about 9:1, about 8:1, about 7:1, about 6:1, about 5:1, about 4:1, about 3:1, about 2:1, about 1:1, about 1:2, about 1:3, about 1:4, about 1:5, about 1:6, about 1:7, about 1:8, about 1:9, about 1:10, about 1:15, about 1:20, or about 1:25; a N77 to Q5216 modification, substitution, and / or substitution ratio of about 25:1, about 20:1, about 15:1, about 10:1, about 9:1, about 8:1, about 7:1, about 6:1, about 5:1, about 4:1, about 3:1, about 2:1, about 1:1, about 1:2, about 1:3, about 1:4, about 1:5, about 1:6, about 1:7, about 1:8, about 1:9, about 1:10, about 1:15, about 1:20, or about 1:25; a modification, substitution, and / or replacement ratio of N77 to N5262 of about 25:1, about 20:1, about 15:1, about 10:1, about 9:1, about 8:1, about 7:1, about 6:1, about 5:1, about 4:1, about 3:1, about 2:1, about 1:1, about 1:2, about 1:3, about 1:4, about 1:5, about 1:6, about 1:7, about 1:8, about 1:9, about 1:10, about 1:15, about 1:20, or about 1:25; an M80 to N93 modification, substitution, and / or replacement ratio of about 25:1, about 20:1, about 15:1, about 10:1, about 9:1, about 8:1, about 7:1, about 6:1, about 5:1, about 4:1, about 3:1, about 2:1, about 1:1, about 1:2, about 1:3, about 1:4, about 1:5, about 1:6, about 1:7, about 1:8, about 1:9, about 1:10, about 1:15, about 1:20, or about 1:25; - a ratio of M80 to M103 modification, substitution, and / or replacement of about 25:1, about 20:1, about 15:1, about 10:1, about 9:1, about 8:1, about 7:1, about 6:1, about 5:1, about 4:1, about 3:1, about 2:1, about 1:1, about 1:2, about 1:3, about 1:4, about 1:5, about 1:6, about 1:7, about 1:8, about 1:9, about 1:10, about 1:15, about 1:20, or about 1:25; - a ratio of M80 to Q125 modification, substitution, and / or replacement of about 25:1, about 20:1, about 15:1, about 10:1, about 9:1, about 8:1, about 7:1, about 6:1, about 5:1, about 4:1, about 3:1, about 2:1, about 1:1, about 1:2, about 1:3, about 1:4, about 1:5, about 1:6, about 1:7, about 1:8, about 1:9, about 1:10, about 1:15, about 1:20, or about 1:25; - a ratio of M80 to N132 modification, substitution, and / or replacement of about 25:1, about 20:1, about 15:1, about 10:1, about 9:1, about 8:1, about 7:1, about 6:1, about 5:1, about 4:1, about 3:1, about 2:1, about 1:1, about 1:2, about 1:3, about 1:4, about 1:5, about 1:6, about 1:7, about 1:8, about 1:9, about 1:10, about 1:15, about 1:20, or about 1:25; - a ratio of M80 to Q139 modifications, substitutions, and / or substitutions of about 25:1, about 20:1, about 15:1, about 10:1, about 9:1, about 8:1, about 7:1, about 6:1, about 5:1, about 4:1, about 3:1, about 2:1, about 1:1, about 1:2, about 1:3, about 1:4, about 1:5, about 1:6, about 1:7, about 1:8, about 1:9, about 1:10, about 1:15, about 1:20, or about 1:25; - a ratio of M80 to Q275 modification, substitution, and / or replacement of about 25:1, about 20:1, about 15:1, about 10:1, about 9:1, about 8:1, about 7:1, about 6:1, about 5:1, about 4:1, about 3:1, about 2:1, about 1:1, about 1:2, about 1:3, about 1:4, about 1:5, about 1:6, about 1:7, about 1:8, about 1:9, about 1:10, about 1:15, about 1:20, or about 1:25; a ratio of M80 to N4191 modification, substitution, and / or replacement of about 25:1, about 20:1, about 15:1, about 10:1, about 9:1, about 8:1, about 7:1, about 6:1, about 5:1, about 4:1, about 3:1, about 2:1, about 1:1, about 1:2, about 1:3, about 1:4, about 1:5, about 1:6, about 1:7, about 1:8, about 1:9, about 1:10, about 1:15, about 1:20, or about 1:25; - a ratio of M80 to Q5216 modifications, substitutions, and / or substitutions of about 25:1, about 20:1, about 15:1, about 10:1, about 9:1, about 8:1, about 7:1, about 6:1, about 5:1, about 4:1, about 3:1, about 2:1, about 1:1, about 1:2, about 1:3, about 1:4, about 1:5, about 1:6, about 1:7, about 1:8, about 1:9, about 1:10, about 1:15, about 1:20, or about 1:25; a ratio of modifications, substitutions, and / or substitutions of M80 to N5262 of about 25:1, about 20:1, about 15:1, about 10:1, about 9:1, about 8:1, about 7:1, about 6:1, about 5:1, about 4:1, about 3:1, about 2:1, about 1:1, about 1:2, about 1:3, about 1:4, about 1:5, about 1:6, about 1:7, about 1:8, about 1:9, about 1:10, about 1:15, about 1:20, or about 1:25; an N93 to M103 modification, substitution, and / or replacement ratio of about 25:1, about 20:1, about 15:1, about 10:1, about 9:1, about 8:1, about 7:1, about 6:1, about 5:1, about 4:1, about 3:1, about 2:1, about 1:1, about 1:2, about 1:3, about 1:4, about 1:5, about 1:6, about 1:7, about 1:8, about 1:9, about 1:10, about 1:15, about 1:20, or about 1:25; an N93 to Q125 modification, substitution, and / or replacement ratio of about 25:1, about 20:1, about 15:1, about 10:1, about 9:1, about 8:1, about 7:1, about 6:1, about 5:1, about 4:1, about 3:1, about 2:1, about 1:1, about 1:2, about 1:3, about 1:4, about 1:5, about 1:6, about 1:7, about 1:8, about 1:9, about 1:10, about 1:15, about 1:20, or about 1:25; an N93 to N132 modification, substitution, and / or replacement ratio of about 25:1, about 20:1, about 15:1, about 10:1, about 9:1, about 8:1, about 7:1, about 6:1, about 5:1, about 4:1, about 3:1, about 2:1, about 1:1, about 1:2, about 1:3, about 1:4, about 1:5, about 1:6, about 1:7, about 1:8, about 1:9, about 1:10, about 1:15, about 1:20, or about 1:25; a N93 to Q139 modification, substitution, and / or replacement ratio of about 25:1, about 20:1, about 15:1, about 10:1, about 9:1, about 8:1, about 7:1, about 6:1, about 5:1, about 4:1, about 3:1, about 2:1, about 1:1, about 1:2, about 1:3, about 1:4, about 1:5, about 1:6, about 1:7, about 1:8, about 1:9, about 1:10, about 1:15, about 1:20, or about 1:25; an N93 to Q275 modification, substitution, and / or replacement ratio of about 25:1, about 20:1, about 15:1, about 10:1, about 9:1, about 8:1, about 7:1, about 6:1, about 5:1, about 4:1, about 3:1, about 2:1, about 1:1, about 1:2, about 1:3, about 1:4, about 1:5, about 1:6, about 1:7, about 1:8, about 1:9, about 1:10, about 1:15, about 1:20, or about 1:25; a modification, substitution, and / or replacement ratio of N93 to N4191 of about 25:1, about 20:1, about 15:1, about 10:1, about 9:1, about 8:1, about 7:1, about 6:1, about 5:1, about 4:1, about 3:1, about 2:1, about 1:1, about 1:2, about 1:3, about 1:4, about 1:5, about 1:6, about 1:7, about 1:8, about 1:9, about 1:10, about 1:15, about 1:20, or about 1:25; - a N93 to Q5216 modification, substitution, and / or replacement ratio of about 25:1, about 20:1, about 15:1, about 10:1, about 9:1, about 8:1, about 7:1, about 6:1, about 5:1, about 4:1, about 3:1, about 2:1, about 1:1, about 1:2, about 1:3, about 1:4, about 1:5, about 1:6, about 1:7, about 1:8, about 1:9, about 1:10, about 1:15, about 1:20, or about 1:25; a modification, substitution, and / or replacement ratio of N93 to N5262 of about 25:1, about 20:1, about 15:1, about 10:1, about 9:1, about 8:1, about 7:1, about 6:1, about 5:1, about 4:1, about 3:1, about 2:1, about 1:1, about 1:2, about 1:3, about 1:4, about 1:5, about 1:6, about 1:7, about 1:8, about 1:9, about 1:10, about 1:15, about 1:20, or about 1:25; - a ratio of M103 to Q125 modification, substitution, and / or replacement of about 25:1, about 20:1, about 15:1, about 10:1, about 9:1, about 8:1, about 7:1, about 6:1, about 5:1, about 4:1, about 3:1, about 2:1, about 1:1, about 1:2, about 1:3, about 1:4, about 1:5, about 1:6, about 1:7, about 1:8, about 1:9, about 1:10, about 1:15, about 1:20, or about 1:25; - a ratio of M103 to N132 modification, substitution, and / or replacement of about 25:1, about 20:1, about 15:1, about 10:1, about 9:1, about 8:1, about 7:1, about 6:1, about 5:1, about 4:1, about 3:1, about 2:1, about 1:1, about 1:2, about 1:3, about 1:4, about 1:5, about 1:6, about 1:7, about 1:8, about 1:9, about 1:10, about 1:15, about 1:20, or about 1:25; - a ratio of M103 to Q139 modifications, substitutions, and / or substitutions of about 25:1, about 20:1, about 15:1, about 10:1, about 9:1, about 8:1, about 7:1, about 6:1, about 5:1, about 4:1, about 3:1, about 2:1, about 1:1, about 1:2, about 1:3, about 1:4, about 1:5, about 1:6, about 1:7, about 1:8, about 1:9, about 1:10, about 1:15, about 1:20, or about 1:25; a ratio of M103 to Q275 modification, substitution, and / or replacement of about 25:1, about 20:1, about 15:1, about 10:1, about 9:1, about 8:1, about 7:1, about 6:1, about 5:1, about 4:1, about 3:1, about 2:1, about 1:1, about 1:2, about 1:3, about 1:4, about 1:5, about 1:6, about 1:7, about 1:8, about 1:9, about 1:10, about 1:15, about 1:20, or about 1:25; a ratio of M103 to N4191 modification, substitution, and / or substitution of about 25:1, about 20:1, about 15:1, about 10:1, about 9:1, about 8:1, about 7:1, about 6:1, about 5:1, about 4:1, about 3:1, about 2:1, about 1:1, about 1:2, about 1:3, about 1:4, about 1:5, about 1:6, about 1:7, about 1:8, about 1:9, about 1:10, about 1:15, about 1:20, or about 1:25; a ratio of M103 to Q5216 modifications, substitutions, and / or substitutions of about 25:1, about 20:1, about 15:1, about 10:1, about 9:1, about 8:1, about 7:1, about 6:1, about 5:1, about 4:1, about 3:1, about 2:1, about 1:1, about 1:2, about 1:3, about 1:4, about 1:5, about 1:6, about 1:7, about 1:8, about 1:9, about 1:10, about 1:15, about 1:20, or about 1:25; a ratio of M103 to N5262 modifications, substitutions, and / or substitutions of about 25:1, about 20:1, about 15:1, about 10:1, about 9:1, about 8:1, about 7:1, about 6:1, about 5:1, about 4:1, about 3:1, about 2:1, about 1:1, about 1:2, about 1:3, about 1:4, about 1:5, about 1:6, about 1:7, about 1:8, about 1:9, about 1:10, about 1:15, about 1:20, or about 1:25; a Q125 to N132 modification, substitution, and / or replacement ratio of about 25:1, about 20:1, about 15:1, about 10:1, about 9:1, about 8:1, about 7:1, about 6:1, about 5:1, about 4:1, about 3:1, about 2:1, about 1:1, about 1:2, about 1:3, about 1:4, about 1:5, about 1:6, about 1:7, about 1:8, about 1:9, about 1:10, about 1:15, about 1:20, or about 1:25; - a ratio of about Q125 to Q139 modification, substitution, and / or replacement of about 25:1, about 20:1, about 15:1, about 10:1, about 9:1, about 8:1, about 7:1, about 6:1, about 5:1, about 4:1, about 3:1, about 2:1, about 1:1, about 1:2, about 1:3, about 1:4, about 1:5, about 1:6, about 1:7, about 1:8, about 1:9, about 1:10, about 1:15, about 1:20, or about 1:25; - a modification, substitution, and / or replacement ratio of about Q125 to Q275 of about 25:1, about 20:1, about 15:1, about 10:1, about 9:1, about 8:1, about 7:1, about 6:1, about 5:1, about 4:1, about 3:1, about 2:1, about 1:1, about 1:2, about 1:3, about 1:4, about 1:5, about 1:6, about 1:7, about 1:8, about 1:9, about 1:10, about 1:15, about 1:20, or about 1:25; a modification, substitution, and / or replacement ratio of Q125 to N4191 of about 25:1, about 20:1, about 15:1, about 10:1, about 9:1, about 8:1, about 7:1, about 6:1, about 5:1, about 4:1, about 3:1, about 2:1, about 1:1, about 1:2, about 1:3, about 1:4, about 1:5, about 1:6, about 1:7, about 1:8, about 1:9, about 1:10, about 1:15, about 1:20, or about 1:25; - a Q125 to Q5216 modification, substitution, and / or substitution ratio of about 25:1, about 20:1, about 15:1, about 10:1, about 9:1, about 8:1, about 7:1, about 6:1, about 5:1, about 4:1, about 3:1, about 2:1, about 1:1, about 1:2, about 1:3, about 1:4, about 1:5, about 1:6, about 1:7, about 1:8, about 1:9, about 1:10, about 1:15, about 1:20, or about 1:25; a modification, substitution, and / or substitution ratio of Q125 to N5262 of about 25:1, about 20:1, about 15:1, about 10:1, about 9:1, about 8:1, about 7:1, about 6:1, about 5:1, about 4:1, about 3:1, about 2:1, about 1:1, about 1:2, about 1:3, about 1:4, about 1:5, about 1:6, about 1:7, about 1:8, about 1:9, about 1:10, about 1:15, about 1:20, or about 1:25; a N132 to Q139 modification, substitution, and / or replacement ratio of about 25:1, about 20:1, about 15:1, about 10:1, about 9:1, about 8:1, about 7:1, about 6:1, about 5:1, about 4:1, about 3:1, about 2:1, about 1:1, about 1:2, about 1:3, about 1:4, about 1:5, about 1:6, about 1:7, about 1:8, about 1:9, about 1:10, about 1:15, about 1:20, or about 1:25; a N132 to Q275 modification, substitution, and / or replacement ratio of about 25:1, about 20:1, about 15:1, about 10:1, about 9:1, about 8:1, about 7:1, about 6:1, about 5:1, about 4:1, about 3:1, about 2:1, about 1:1, about 1:2, about 1:3, about 1:4, about 1:5, about 1:6, about 1:7, about 1:8, about 1:9, about 1:10, about 1:15, about 1:20, or about 1:25; a modification, substitution, and / or replacement ratio of N132 to N4191 of about 25:1, about 20:1, about 15:1, about 10:1, about 9:1, about 8:1, about 7:1, about 6:1, about 5:1, about 4:1, about 3:1, about 2:1, about 1:1, about 1:2, about 1:3, about 1:4, about 1:5, about 1:6, about 1:7, about 1:8, about 1:9, about 1:10, about 1:15, about 1:20, or about 1:25; - a N132 to Q5216 modification, substitution, and / or substitution ratio of about 25:1, about 20:1, about 15:1, about 10:1, about 9:1, about 8:1, about 7:1, about 6:1, about 5:1, about 4:1, about 3:1, about 2:1, about 1:1, about 1:2, about 1:3, about 1:4, about 1:5, about 1:6, about 1:7, about 1:8, about 1:9, about 1:10, about 1:15, about 1:20, or about 1:25; a modification, substitution, and / or replacement ratio of N132 to N5262 of about 25:1, about 20:1, about 15:1, about 10:1, about 9:1, about 8:1, about 7:1, about 6:1, about 5:1, about 4:1, about 3:1, about 2:1, about 1:1, about 1:2, about 1:3, about 1:4, about 1:5, about 1:6, about 1:7, about 1:8, about 1:9, about 1:10, about 1:15, about 1:20, or about 1:25; a Q139 to Q275 modification, substitution, and / or replacement ratio of about 25:1, about 20:1, about 15:1, about 10:1, about 9:1, about 8:1, about 7:1, about 6:1, about 5:1, about 4:1, about 3:1, about 2:1, about 1:1, about 1:2, about 1:3, about 1:4, about 1:5, about 1:6, about 1:7, about 1:8, about 1:9, about 1:10, about 1:15, about 1:20, or about 1:25; a modification, substitution, and / or replacement ratio of Q139 to N4191 of about 25:1, about 20:1, about 15:1, about 10:1, about 9:1, about 8:1, about 7:1, about 6:1, about 5:1, about 4:1, about 3:1, about 2:1, about 1:1, about 1:2, about 1:3, about 1:4, about 1:5, about 1:6, about 1:7, about 1:8, about 1:9, about 1:10, about 1:15, about 1:20, or about 1:25; a Q139 to Q5216 modification, substitution, and / or substitution ratio of about 25:1, about 20:1, about 15:1, about 10:1, about 9:1, about 8:1, about 7:1, about 6:1, about 5:1, about 4:1, about 3:1, about 2:1, about 1:1, about 1:2, about 1:3, about 1:4, about 1:5, about 1:6, about 1:7, about 1:8, about 1:9, about 1:10, about 1:15, about 1:20, or about 1:25; a modification, substitution, and / or replacement ratio of Q139 to N5262 of about 25:1, about 20:1, about 15:1, about 10:1, about 9:1, about 8:1, about 7:1, about 6:1, about 5:1, about 4:1, about 3:1, about 2:1, about 1:1, about 1:2, about 1:3, about 1:4, about 1:5, about 1:6, about 1:7, about 1:8, about 1:9, about 1:10, about 1:15, about 1:20, or about 1:25; a modification, substitution, and / or replacement ratio of Q275 to N4191 of about 25:1, about 20:1, about 15:1, about 10:1, about 9:1, about 8:1, about 7:1, about 6:1, about 5:1, about 4:1, about 3:1, about 2:1, about 1:1, about 1:2, about 1:3, about 1:4, about 1:5, about 1:6, about 1:7, about 1:8, about 1:9, about 1:10, about 1:15, about 1:20, or about 1:25; a modification, substitution, and / or substitution ratio of Q275 to Q5216 of about 25:1, about 20:1, about 15:1, about 10:1, about 9:1, about 8:1, about 7:1, about 6:1, about 5:1, about 4:1, about 3:1, about 2:1, about 1:1, about 1:2, about 1:3, about 1:4, about 1:5, about 1:6, about 1:7, about 1:8, about 1:9, about 1:10, about 1:15, about 1:20, or about 1:25 a modification, substitution, and / or replacement ratio of Q275 to N5262 of about 25:1, about 20:1, about 15:1, about 10:1, about 9:1, about 8:1, about 7:1, about 6:1, about 5:1, about 4:1, about 3:1, about 2:1, about 1:1, about 1:2, about 1:3, about 1:4, about 1:5, about 1:6, about 1:7, about 1:8, about 1:9, about 1:10, about 1:15, about 1:20, or about 1:25; - a modification, substitution, and / or substitution ratio of N4191 to Q5216 of about 25:1, about 20:1, about 15:1, about 10:1, about 9:1, about 8:1, about 7:1, about 6:1, about 5:1, about 4:1, about 3:1, about 2:1, about 1:1, about 1:2, about 1:3, about 1:4, about 1:5, about 1:6, about 1:7, about 1:8, about 1:9, about 1:10, about 1:15, about 1:20, or about 1:25; a modification, substitution, and / or replacement ratio of N4191 to N5262 of about 25:1, about 20:1, about 15:1, about 10:1, about 9:1, about 8:1, about 7:1, about 6:1, about 5:1, about 4:1, about 3:1, about 2:1, about 1:1, about 1:2, about 1:3, about 1:4, about 1:5, about 1:6, about 1:7, about 1:8, about 1:9, about 1:10, about 1:15, about 1:20, or about 1:25; a modification, substitution, and / or substitution ratio of Q5216 to N5262 of about 25:1, about 20:1, about 15:1, about 10:1, about 9:1, about 8:1, about 7:1, about 6:1, about 5:1, about 4:1, about 3:1, about 2:1, about 1:1, about 1:2, about 1:3, about 1:4, about 1:5, about 1:6, about 1:7, about 1:8, about 1:9, about 1:10, about 1:15, about 1:20, or about 1:25;

[0009] In some embodiments, the present disclosure provides compositions that include a plurality of peptides or protein fragments, e.g., a plurality of fibroin heavy and light chain peptides or fibroin heavy and light chain fragments, and that include a plurality of amino acid modifications, substitutions, and / or substitutions, and further include, at specific positions, ratios of one or more modifications, substitutions, and / or substitutions selected from the following: - a ratio of heavy chain Q58 to light chain N23, Q24, N28, M69, N105, N108, N118, N136, N138, Q149, N186, N200, Q202, N204, N240, N248 or Q255 modifications, substitutions and / or substitutions of about 25:1, about 20:1, about 15:1, about 10:1, about 9:1, about 8:1, about 7:1, about 6:1, about 5:1, about 4:1, about 3:1, about 2:1, about 1:1, about 1:2, about 1:3, about 1:4, about 1:5, about 1:6, about 1:7, about 1:8, about 1:9, about 1:10, about 1:15, about 1:20, or about 1:25; - a ratio of heavy chain M64 to light chain N23, Q24, N28, M69, N105, N108, N118, N136, N138, Q149, N186, N200, Q202, N204, N240, N248, or Q255 modifications, substitutions, and / or substitutions of about 25:1, about 20:1, about 15:1, about 10:1, about 9:1, about 8:1, about 7:1, about 6:1, about 5:1, about 4:1, about 3:1, about 2:1, about 1:1, about 1:2, about 1:3, about 1:4, about 1:5, about 1:6, about 1:7, about 1:8, about 1:9, about 1:10, about 1:15, about 1:20, or about 1:25; - a ratio of heavy chain N68 to light chain N23, Q24, N28, M69, N105, N108, N118, N136, N138, Q149, N186, N200, Q202, N204, N240, N248 or Q255 modifications, substitutions, and / or substitutions of about 25:1, about 20:1, about 15:1, about 10:1, about 9:1, about 8:1, about 7:1, about 6:1, about 5:1, about 4:1, about 3:1, about 2:1, about 1:1, about 1:2, about 1:3, about 1:4, about 1:5, about 1:6, about 1:7, about 1:8, about 1:9, about 1:10, about 1:15, about 1:20, or about 1:25; - a ratio of heavy chain N70 to light chain N23, Q24, N28, M69, N105, N108, N118, N136, N138, Q149, N186, N200, Q202, N204, N240, N248 or Q255 modifications, substitutions, and / or substitutions of about 25:1, about 20:1, about 15:1, about 10:1, about 9:1, about 8:1, about 7:1, about 6:1, about 5:1, about 4:1, about 3:1, about 2:1, about 1:1, about 1:2, about 1:3, about 1:4, about 1:5, about 1:6, about 1:7, about 1:8, about 1:9, about 1:10, about 1:15, about 1:20, or about 1:25; - a ratio of heavy chain N77 to light chain N23, Q24, N28, M69, N105, N108, N118, N136, N138, Q149, N186, N200, Q202, N204, N240, N248 or Q255 modifications, substitutions, and / or substitutions of about 25:1, about 20:1, about 15:1, about 10:1, about 9:1, about 8:1, about 7:1, about 6:1, about 5:1, about 4:1, about 3:1, about 2:1, about 1:1, about 1:2, about 1:3, about 1:4, about 1:5, about 1:6, about 1:7, about 1:8, about 1:9, about 1:10, about 1:15, about 1:20, or about 1:25; - a ratio of heavy chain M80 to light chain N23, Q24, N28, M69, N105, N108, N118, N136, N138, Q149, N186, N200, Q202, N204, N240, N248 or Q255 modifications, substitutions, and / or substitutions of about 25:1, about 20:1, about 15:1, about 10:1, about 9:1, about 8:1, about 7:1, about 6:1, about 5:1, about 4:1, about 3:1, about 2:1, about 1:1, about 1:2, about 1:3, about 1:4, about 1:5, about 1:6, about 1:7, about 1:8, about 1:9, about 1:10, about 1:15, about 1:20, or about 1:25; - a ratio of heavy chain N93 to light chain N23, Q24, N28, M69, N105, N108, N118, N136, N138, Q149, N186, N200, Q202, N204, N240, N248 or Q255 modifications, substitutions, and / or substitutions of about 25:1, about 20:1, about 15:1, about 10:1, about 9:1, about 8:1, about 7:1, about 6:1, about 5:1, about 4:1, about 3:1, about 2:1, about 1:1, about 1:2, about 1:3, about 1:4, about 1:5, about 1:6, about 1:7, about 1:8, about 1:9, about 1:10, about 1:15, about 1:20, or about 1:25; - a ratio of heavy chain M103 to light chain N23, Q24, N28, M69, N105, N108, N118, N136, N138, Q149, N186, N200, Q202, N204, N240, N248 or Q255 modifications, substitutions, and / or substitutions of about 25:1, about 20:1, about 15:1, about 10:1, about 9:1, about 8:1, about 7:1, about 6:1, about 5:1, about 4:1, about 3:1, about 2:1, about 1:1, about 1:2, about 1:3, about 1:4, about 1:5, about 1:6, about 1:7, about 1:8, about 1:9, about 1:10, about 1:15, about 1:20, or about 1:25; - a ratio of heavy chain Q125 to light chain N23, Q24, N28, M69, N105, N108, N118, N136, N138, Q149, N186, N200, Q202, N204, N240, N248 or Q255 modifications, substitutions, and / or substitutions of about 25:1, about 20:1, about 15:1, about 10:1, about 9:1, about 8:1, about 7:1, about 6:1, about 5:1, about 4:1, about 3:1, about 2:1, about 1:1, about 1:2, about 1:3, about 1:4, about 1:5, about 1:6, about 1:7, about 1:8, about 1:9, about 1:10, about 1:15, about 1:20, or about 1:25; - a ratio of heavy chain N132 to light chain N23, Q24, N28, M69, N105, N108, N118, N136, N138, Q149, N186, N200, Q202, N204, N240, N248 or Q255 modifications, substitutions and / or substitutions of about 25:1, about 20:1, about 15:1, about 10:1, about 9:1, about 8:1, about 7:1, about 6:1, about 5:1, about 4:1, about 3:1, about 2:1, about 1:1, about 1:2, about 1:3, about 1:4, about 1:5, about 1:6, about 1:7, about 1:8, about 1:9, about 1:10, about 1:15, about 1:20, or about 1:25; - a ratio of heavy chain Q139 to light chain N23, Q24, N28, M69, N105, N108, N118, N136, N138, Q149, N186, N200, Q202, N204, N240, N248 or Q255 modifications, substitutions, and / or substitutions of about 25:1, about 20:1, about 15:1, about 10:1, about 9:1, about 8:1, about 7:1, about 6:1, about 5:1, about 4:1, about 3:1, about 2:1, about 1:1, about 1:2, about 1:3, about 1:4, about 1:5, about 1:6, about 1:7, about 1:8, about 1:9, about 1:10, about 1:15, about 1:20, or about 1:25; - a ratio of heavy chain Q275 to light chain N23, Q24, N28, M69, N105, N108, N118, N136, N138, Q149, N186, N200, Q202, N204, N240, N248 or Q255 modifications, substitutions, and / or substitutions of about 25:1, about 20:1, about 15:1, about 10:1, about 9:1, about 8:1, about 7:1, about 6:1, about 5:1, about 4:1, about 3:1, about 2:1, about 1:1, about 1:2, about 1:3, about 1:4, about 1:5, about 1:6, about 1:7, about 1:8, about 1:9, about 1:10, about 1:15, about 1:20, or about 1:25; - a ratio of heavy chain N4191 to light chain N23, Q24, N28, M69, N105, N108, N118, N136, N138, Q149, N186, N200, Q202, N204, N240, N248 or Q255 modifications, substitutions, and / or substitutions of about 25:1, about 20:1, about 15:1, about 10:1, about 9:1, about 8:1, about 7:1, about 6:1, about 5:1, about 4:1, about 3:1, about 2:1, about 1:1, about 1:2, about 1:3, about 1:4, about 1:5, about 1:6, about 1:7, about 1:8, about 1:9, about 1:10, about 1:15, about 1:20, or about 1:25; - a ratio of heavy chain Q5216 to light chain N23, Q24, N28, M69, N105, N108, N118, N136, N138, Q149, N186, N200, Q202, N204, N240, N248 or Q255 modifications, substitutions, and / or substitutions of about 25:1, about 20:1, about 15:1, about 10:1, about 9:1, about 8:1, about 7:1, about 6:1, about 5:1, about 4:1, about 3:1, about 2:1, about 1:1, about 1:2, about 1:3, about 1:4, about 1:5, about 1:6, about 1:7, about 1:8, about 1:9, about 1:10, about 1:15, about 1:20, or about 1:25; - a ratio of heavy chain N5262 to light chain N23, Q24, N28, M69, N105, N108, N118, N136, N138, Q149, N186, N200, Q202, N204, N240, N248 or Q255 modifications, substitutions, and / or substitutions of about 25:1, about 20:1, about 15:1, about 10:1, about 9:1, about 8:1, about 7:1, about 6:1, about 5:1, about 4:1, about 3:1, about 2:1, about 1:1, about 1:2, about 1:3, about 1:4, about 1:5, about 1:6, about 1:7, about 1:8, about 1:9, about 1:10, about 1:15, about 1:20, or about 1:25;

[0010] In some embodiments, the present disclosure provides compositions comprising a plurality of peptides or protein fragments, e.g., a plurality of fibroin heavy chain and p25 peptides or fibroin heavy chain and p25 fragments, and comprising a plurality of amino acid modifications, substitutions, and / or substitutions, wherein specific positions comprise a ratio of one or more modifications, substitutions, and / or substitutions selected from the following: - a ratio of heavy chain Q58 to p25 Q62, N93, M120, N149, N172, N174, or N202 modifications, substitutions, and / or substitutions of about 25:1, about 20:1, about 15:1, about 10:1, about 9:1, about 8:1, about 7:1, about 6:1, about 5:1, about 4:1, about 3:1, about 2:1, about 1:1, about 1:2, about 1:3, about 1:4, about 1:5, about 1:6, about 1:7, about 1:8, about 1:9, about 1:10, about 1:15, about 1:20, or about 1:25; - a ratio of heavy chain M64 to p25 Q62, N93, M120, N149, N172, N174, or N202 modifications, substitutions, and / or substitutions of about 25:1, about 20:1, about 15:1, about 10:1, about 9:1, about 8:1, about 7:1, about 6:1, about 5:1, about 4:1, about 3:1, about 2:1, about 1:1, about 1:2, about 1:3, about 1:4, about 1:5, about 1:6, about 1:7, about 1:8, about 1:9, about 1:10, about 1:15, about 1:20, or about 1:25; - a ratio of heavy chain N68 to p25 Q62, N93, M120, N149, N172, N174, or N202 modifications, substitutions, and / or substitutions of about 25:1, about 20:1, about 15:1, about 10:1, about 9:1, about 8:1, about 7:1, about 6:1, about 5:1, about 4:1, about 3:1, about 2:1, about 1:1, about 1:2, about 1:3, about 1:4, about 1:5, about 1:6, about 1:7, about 1:8, about 1:9, about 1:10, about 1:15, about 1:20, or about 1:25; - a ratio of heavy chain N70 to p25 Q62, N93, M120, N149, N172, N174, or N202 modifications, substitutions, and / or substitutions of about 25:1, about 20:1, about 15:1, about 10:1, about 9:1, about 8:1, about 7:1, about 6:1, about 5:1, about 4:1, about 3:1, about 2:1, about 1:1, about 1:2, about 1:3, about 1:4, about 1:5, about 1:6, about 1:7, about 1:8, about 1:9, about 1:10, about 1:15, about 1:20, or about 1:25; - a ratio of heavy chain N77 to p25 Q62, N93, M120, N149, N172, N174, or N202 modifications, substitutions, and / or substitutions of about 25:1, about 20:1, about 15:1, about 10:1, about 9:1, about 8:1, about 7:1, about 6:1, about 5:1, about 4:1, about 3:1, about 2:1, about 1:1, about 1:2, about 1:3, about 1:4, about 1:5, about 1:6, about 1:7, about 1:8, about 1:9, about 1:10, about 1:15, about 1:20, or about 1:25; - a ratio of heavy chain M80 to p25 Q62, N93, M120, N149, N172, N174, or N202 modifications, substitutions, and / or substitutions of about 25:1, about 20:1, about 15:1, about 10:1, about 9:1, about 8:1, about 7:1, about 6:1, about 5:1, about 4:1, about 3:1, about 2:1, about 1:1, about 1:2, about 1:3, about 1:4, about 1:5, about 1:6, about 1:7, about 1:8, about 1:9, about 1:10, about 1:15, about 1:20, or about 1:25; - a ratio of heavy chain N93 to p25 Q62, N93, M120, N149, N172, N174, or N202 modifications, substitutions, and / or substitutions of about 25:1, about 20:1, about 15:1, about 10:1, about 9:1, about 8:1, about 7:1, about 6:1, about 5:1, about 4:1, about 3:1, about 2:1, about 1:1, about 1:2, about 1:3, about 1:4, about 1:5, about 1:6, about 1:7, about 1:8, about 1:9, about 1:10, about 1:15, about 1:20, or about 1:25; - a ratio of heavy chain M103 to p25 Q62, N93, M120, N149, N172, N174, or N202 modifications, substitutions, and / or substitutions of about 25:1, about 20:1, about 15:1, about 10:1, about 9:1, about 8:1, about 7:1, about 6:1, about 5:1, about 4:1, about 3:1, about 2:1, about 1:1, about 1:2, about 1:3, about 1:4, about 1:5, about 1:6, about 1:7, about 1:8, about 1:9, about 1:10, about 1:15, about 1:20, or about 1:25; - a ratio of heavy chain Q125 to p25 Q62, N93, M120, N149, N172, N174, or N202 modifications, substitutions, and / or substitutions of about 25:1, about 20:1, about 15:1, about 10:1, about 9:1, about 8:1, about 7:1, about 6:1, about 5:1, about 4:1, about 3:1, about 2:1, about 1:1, about 1:2, about 1:3, about 1:4, about 1:5, about 1:6, about 1:7, about 1:8, about 1:9, about 1:10, about 1:15, about 1:20, or about 1:25; - a ratio of heavy chain N132 to p25 Q62, N93, M120, N149, N172, N174, or N202 modifications, substitutions, and / or substitutions of about 25:1, about 20:1, about 15:1, about 10:1, about 9:1, about 8:1, about 7:1, about 6:1, about 5:1, about 4:1, about 3:1, about 2:1, about 1:1, about 1:2, about 1:3, about 1:4, about 1:5, about 1:6, about 1:7, about 1:8, about 1:9, about 1:10, about 1:15, about 1:20, or about 1:25; - a ratio of heavy chain Q139 to p25 Q62, N93, M120, N149, N172, N174, or N202 modifications, substitutions, and / or substitutions of about 25:1, about 20:1, about 15:1, about 10:1, about 9:1, about 8:1, about 7:1, about 6:1, about 5:1, about 4:1, about 3:1, about 2:1, about 1:1, about 1:2, about 1:3, about 1:4, about 1:5, about 1:6, about 1:7, about 1:8, about 1:9, about 1:10, about 1:15, about 1:20, or about 1:25; - a ratio of heavy chain Q275 to p25 Q62, N93, M120, N149, N172, N174, or N202 modifications, substitutions, and / or substitutions of about 25:1, about 20:1, about 15:1, about 10:1, about 9:1, about 8:1, about 7:1, about 6:1, about 5:1, about 4:1, about 3:1, about 2:1, about 1:1, about 1:2, about 1:3, about 1:4, about 1:5, about 1:6, about 1:7, about 1:8, about 1:9, about 1:10, about 1:15, about 1:20, or about 1:25; - a ratio of heavy chain N4191 to p25 Q62, N93, M120, N149, N172, N174, or N202 modifications, substitutions, and / or substitutions of about 25:1, about 20:1, about 15:1, about 10:1, about 9:1, about 8:1, about 7:1, about 6:1, about 5:1, about 4:1, about 3:1, about 2:1, about 1:1, about 1:2, about 1:3, about 1:4, about 1:5, about 1:6, about 1:7, about 1:8, about 1:9, about 1:10, about 1:15, about 1:20, or about 1:25; - a ratio of heavy chain Q5216 to p25 Q62, N93, M120, N149, N172, N174, or N202 modifications, substitutions, and / or substitutions of about 25:1, about 20:1, about 15:1, about 10:1, about 9:1, about 8:1, about 7:1, about 6:1, about 5:1, about 4:1, about 3:1, about 2:1, about 1:1, about 1:2, about 1:3, about 1:4, about 1:5, about 1:6, about 1:7, about 1:8, about 1:9, about 1:10, about 1:15, about 1:20, or about 1:25; - a ratio of heavy chain N5262 to p25 Q62, N93, M120, N149, N172, N174, or N202 modifications, substitutions, and / or substitutions of about 25:1, about 20:1, about 15:1, about 10:1, about 9:1, about 8:1, about 7:1, about 6:1, about 5:1, about 4:1, about 3:1, about 2:1, about 1:1, about 1:2, about 1:3, about 1:4, about 1:5, about 1:6, about 1:7, about 1:8, about 1:9, about 1:10, about 1:15, about 1:20, or about 1:25;

[0011] In some embodiments, the present disclosure provides compositions comprising a plurality of peptides or protein fragments, e.g., a plurality of light chain fibroin peptides or fibroin fragments, and comprising a plurality of amino acid modifications, substitutions, and / or substitutions, wherein specific positions comprise a ratio of one or more modifications, substitutions, and / or substitutions selected from the following: an N23 to Q24 modification, substitution, and / or replacement ratio of about 25:1, about 20:1, about 15:1, about 10:1, about 9:1, about 8:1, about 7:1, about 6:1, about 5:1, about 4:1, about 3:1, about 2:1, about 1:1, about 1:2, about 1:3, about 1:4, about 1:5, about 1:6, about 1:7, about 1:8, about 1:9, about 1:10, about 1:15, about 1:20, or about 1:25; a Q24 to N28 modification, substitution, and / or replacement ratio of about 25:1, about 20:1, about 15:1, about 10:1, about 9:1, about 8:1, about 7:1, about 6:1, about 5:1, about 4:1, about 3:1, about 2:1, about 1:1, about 1:2, about 1:3, about 1:4, about 1:5, about 1:6, about 1:7, about 1:8, about 1:9, about 1:10, about 1:15, about 1:20, or about 1:25; a Q24 to M69 modification, substitution, and / or replacement ratio of about 25:1, about 20:1, about 15:1, about 10:1, about 9:1, about 8:1, about 7:1, about 6:1, about 5:1, about 4:1, about 3:1, about 2:1, about 1:1, about 1:2, about 1:3, about 1:4, about 1:5, about 1:6, about 1:7, about 1:8, about 1:9, about 1:10, about 1:15, about 1:20, or about 1:25; a Q24 to N105 modification, substitution, and / or replacement ratio of about 25:1, about 20:1, about 15:1, about 10:1, about 9:1, about 8:1, about 7:1, about 6:1, about 5:1, about 4:1, about 3:1, about 2:1, about 1:1, about 1:2, about 1:3, about 1:4, about 1:5, about 1:6, about 1:7, about 1:8, about 1:9, about 1:10, about 1:15, about 1:20, or about 1:25; a Q24 to N108 modification, substitution, and / or replacement ratio of about 25:1, about 20:1, about 15:1, about 10:1, about 9:1, about 8:1, about 7:1, about 6:1, about 5:1, about 4:1, about 3:1, about 2:1, about 1:1, about 1:2, about 1:3, about 1:4, about 1:5, about 1:6, about 1:7, about 1:8, about 1:9, about 1:10, about 1:15, about 1:20, or about 1:25; a Q24 to N118 modification, substitution, and / or replacement ratio of about 25:1, about 20:1, about 15:1, about 10:1, about 9:1, about 8:1, about 7:1, about 6:1, about 5:1, about 4:1, about 3:1, about 2:1, about 1:1, about 1:2, about 1:3, about 1:4, about 1:5, about 1:6, about 1:7, about 1:8, about 1:9, about 1:10, about 1:15, about 1:20, or about 1:25; a Q24 to N136 modification, substitution, and / or replacement ratio of about 25:1, about 20:1, about 15:1, about 10:1, about 9:1, about 8:1, about 7:1, about 6:1, about 5:1, about 4:1, about 3:1, about 2:1, about 1:1, about 1:2, about 1:3, about 1:4, about 1:5, about 1:6, about 1:7, about 1:8, about 1:9, about 1:10, about 1:15, about 1:20, or about 1:25; a Q24 to N138 modification, substitution, and / or replacement ratio of about 25:1, about 20:1, about 15:1, about 10:1, about 9:1, about 8:1, about 7:1, about 6:1, about 5:1, about 4:1, about 3:1, about 2:1, about 1:1, about 1:2, about 1:3, about 1:4, about 1:5, about 1:6, about 1:7, about 1:8, about 1:9, about 1:10, about 1:15, about 1:20, or about 1:25; a Q24 to Q149 modification, substitution, and / or replacement ratio of about 25:1, about 20:1, about 15:1, about 10:1, about 9:1, about 8:1, about 7:1, about 6:1, about 5:1, about 4:1, about 3:1, about 2:1, about 1:1, about 1:2, about 1:3, about 1:4, about 1:5, about 1:6, about 1:7, about 1:8, about 1:9, about 1:10, about 1:15, about 1:20, or about 1:25; a Q24 to N186 modification, substitution, and / or replacement ratio of about 25:1, about 20:1, about 15:1, about 10:1, about 9:1, about 8:1, about 7:1, about 6:1, about 5:1, about 4:1, about 3:1, about 2:1, about 1:1, about 1:2, about 1:3, about 1:4, about 1:5, about 1:6, about 1:7, about 1:8, about 1:9, about 1:10, about 1:15, about 1:20, or about 1:25; a Q24 to N200 modification, substitution, and / or replacement ratio of about 25:1, about 20:1, about 15:1, about 10:1, about 9:1, about 8:1, about 7:1, about 6:1, about 5:1, about 4:1, about 3:1, about 2:1, about 1:1, about 1:2, about 1:3, about 1:4, about 1:5, about 1:6, about 1:7, about 1:8, about 1:9, about 1:10, about 1:15, about 1:20, or about 1:25; a modification, substitution, and / or substitution ratio of Q24 to Q202 of about 25:1, about 20:1, about 15:1, about 10:1, about 9:1, about 8:1, about 7:1, about 6:1, about 5:1, about 4:1, about 3:1, about 2:1, about 1:1, about 1:2, about 1:3, about 1:4, about 1:5, about 1:6, about 1:7, about 1:8, about 1:9, about 1:10, about 1:15, about 1:20, or about 1:25; a Q24 to N204 modification, substitution, and / or replacement ratio of about 25:1, about 20:1, about 15:1, about 10:1, about 9:1, about 8:1, about 7:1, about 6:1, about 5:1, about 4:1, about 3:1, about 2:1, about 1:1, about 1:2, about 1:3, about 1:4, about 1:5, about 1:6, about 1:7, about 1:8, about 1:9, about 1:10, about 1:15, about 1:20, or about 1:25; a Q24 to N240 modification, substitution, and / or replacement ratio of about 25:1, about 20:1, about 15:1, about 10:1, about 9:1, about 8:1, about 7:1, about 6:1, about 5:1, about 4:1, about 3:1, about 2:1, about 1:1, about 1:2, about 1:3, about 1:4, about 1:5, about 1:6, about 1:7, about 1:8, about 1:9, about 1:10, about 1:15, about 1:20, or about 1:25; a Q24 to N248 modification, substitution, and / or replacement ratio of about 25:1, about 20:1, about 15:1, about 10:1, about 9:1, about 8:1, about 7:1, about 6:1, about 5:1, about 4:1, about 3:1, about 2:1, about 1:1, about 1:2, about 1:3, about 1:4, about 1:5, about 1:6, about 1:7, about 1:8, about 1:9, about 1:10, about 1:15, about 1:20, or about 1:25; a Q24 to Q255 modification, substitution, and / or substitution ratio of about 25:1, about 20:1, about 15:1, about 10:1, about 9:1, about 8:1, about 7:1, about 6:1, about 5:1, about 4:1, about 3:1, about 2:1, about 1:1, about 1:2, about 1:3, about 1:4, about 1:5, about 1:6, about 1:7, about 1:8, about 1:9, about 1:10, about 1:15, about 1:20, or about 1:25; an N28 to M69 modification, substitution, and / or replacement ratio of about 25:1, about 20:1, about 15:1, about 10:1, about 9:1, about 8:1, about 7:1, about 6:1, about 5:1, about 4:1, about 3:1, about 2:1, about 1:1, about 1:2, about 1:3, about 1:4, about 1:5, about 1:6, about 1:7, about 1:8, about 1:9, about 1:10, about 1:15, about 1:20, or about 1:25; a N28 to N105 modification, substitution, and / or replacement ratio of about 25:1, about 20:1, about 15:1, about 10:1, about 9:1, about 8:1, about 7:1, about 6:1, about 5:1, about 4:1, about 3:1, about 2:1, about 1:1, about 1:2, about 1:3, about 1:4, about 1:5, about 1:6, about 1:7, about 1:8, about 1:9, about 1:10, about 1:15, about 1:20, or about 1:25; - a N28 to N108 modification, substitution, and / or replacement ratio of about 25:1, about 20:1, about 15:1, about 10:1, about 9:1, about 8:1, about 7:1, about 6:1, about 5:1, about 4:1, about 3:1, about 2:1, about 1:1, about 1:2, about 1:3, about 1:4, about 1:5, about 1:6, about 1:7, about 1:8, about 1:9, about 1:10, about 1:15, about 1:20, or about 1:25; - a N28 to N118 modification, substitution, and / or replacement ratio of about 25:1, about 20:1, about 15:1, about 10:1, about 9:1, about 8:1, about 7:1, about 6:1, about 5:1, about 4:1, about 3:1, about 2:1, about 1:1, about 1:2, about 1:3, about 1:4, about 1:5, about 1:6, about 1:7, about 1:8, about 1:9, about 1:10, about 1:15, about 1:20, or about 1:25; a N28 to N136 modification, substitution, and / or replacement ratio of about 25:1, about 20:1, about 15:1, about 10:1, about 9:1, about 8:1, about 7:1, about 6:1, about 5:1, about 4:1, about 3:1, about 2:1, about 1:1, about 1:2, about 1:3, about 1:4, about 1:5, about 1:6, about 1:7, about 1:8, about 1:9, about 1:10, about 1:15, about 1:20, or about 1:25; a N28 to N138 modification, substitution, and / or replacement ratio of about 25:1, about 20:1, about 15:1, about 10:1, about 9:1, about 8:1, about 7:1, about 6:1, about 5:1, about 4:1, about 3:1, about 2:1, about 1:1, about 1:2, about 1:3, about 1:4, about 1:5, about 1:6, about 1:7, about 1:8, about 1:9, about 1:10, about 1:15, about 1:20, or about 1:25; a N28 to Q149 modification, substitution, and / or replacement ratio of about 25:1, about 20:1, about 15:1, about 10:1, about 9:1, about 8:1, about 7:1, about 6:1, about 5:1, about 4:1, about 3:1, about 2:1, about 1:1, about 1:2, about 1:3, about 1:4, about 1:5, about 1:6, about 1:7, about 1:8, about 1:9, about 1:10, about 1:15, about 1:20, or about 1:25; - a N28 to N186 modification, substitution, and / or replacement ratio of about 25:1, about 20:1, about 15:1, about 10:1, about 9:1, about 8:1, about 7:1, about 6:1, about 5:1, about 4:1, about 3:1, about 2:1, about 1:1, about 1:2, about 1:3, about 1:4, about 1:5, about 1:6, about 1:7, about 1:8, about 1:9, about 1:10, about 1:15, about 1:20, or about 1:25; - a N28 to N200 modification, substitution, and / or replacement ratio of about 25:1, about 20:1, about 15:1, about 10:1, about 9:1, about 8:1, about 7:1, about 6:1, about 5:1, about 4:1, about 3:1, about 2:1, about 1:1, about 1:2, about 1:3, about 1:4, about 1:5, about 1:6, about 1:7, about 1:8, about 1:9, about 1:10, about 1:15, about 1:20, or about 1:25; a N28 to Q202 modification, substitution, and / or substitution ratio of about 25:1, about 20:1, about 15:1, about 10:1, about 9:1, about 8:1, about 7:1, about 6:1, about 5:1, about 4:1, about 3:1, about 2:1, about 1:1, about 1:2, about 1:3, about 1:4, about 1:5, about 1:6, about 1:7, about 1:8, about 1:9, about 1:10, about 1:15, about 1:20, or about 1:25; a N28 to N204 modification, substitution, and / or replacement ratio of about 25:1, about 20:1, about 15:1, about 10:1, about 9:1, about 8:1, about 7:1, about 6:1, about 5:1, about 4:1, about 3:1, about 2:1, about 1:1, about 1:2, about 1:3, about 1:4, about 1:5, about 1:6, about 1:7, about 1:8, about 1:9, about 1:10, about 1:15, about 1:20, or about 1:25; - a N28 to N240 modification, substitution, and / or replacement ratio of about 25:1, about 20:1, about 15:1, about 10:1, about 9:1, about 8:1, about 7:1, about 6:1, about 5:1, about 4:1, about 3:1, about 2:1, about 1:1, about 1:2, about 1:3, about 1:4, about 1:5, about 1:6, about 1:7, about 1:8, about 1:9, about 1:10, about 1:15, about 1:20, or about 1:25; a N28 to N248 modification, substitution, and / or replacement ratio of about 25:1, about 20:1, about 15:1, about 10:1, about 9:1, about 8:1, about 7:1, about 6:1, about 5:1, about 4:1, about 3:1, about 2:1, about 1:1, about 1:2, about 1:3, about 1:4, about 1:5, about 1:6, about 1:7, about 1:8, about 1:9, about 1:10, about 1:15, about 1:20, or about 1:25; a N28 to Q255 modification, substitution, and / or replacement ratio of about 25:1, about 20:1, about 15:1, about 10:1, about 9:1, about 8:1, about 7:1, about 6:1, about 5:1, about 4:1, about 3:1, about 2:1, about 1:1, about 1:2, about 1:3, about 1:4, about 1:5, about 1:6, about 1:7, about 1:8, about 1:9, about 1:10, about 1:15, about 1:20, or about 1:25; - an M69 to N105 modification, substitution, and / or replacement ratio of about 25:1, about 20:1, about 15:1, about 10:1, about 9:1, about 8:1, about 7:1, about 6:1, about 5:1, about 4:1, about 3:1, about 2:1, about 1:1, about 1:2, about 1:3, about 1:4, about 1:5, about 1:6, about 1:7, about 1:8, about 1:9, about 1:10, about 1:15, about 1:20, or about 1:25; - an M69 to N108 modification, substitution, and / or replacement ratio of about 25:1, about 20:1, about 15:1, about 10:1, about 9:1, about 8:1, about 7:1, about 6:1, about 5:1, about 4:1, about 3:1, about 2:1, about 1:1, about 1:2, about 1:3, about 1:4, about 1:5, about 1:6, about 1:7, about 1:8, about 1:9, about 1:10, about 1:15, about 1:20, or about 1:25; - an M69 to N118 modification, substitution, and / or replacement ratio of about 25:1, about 20:1, about 15:1, about 10:1, about 9:1, about 8:1, about 7:1, about 6:1, about 5:1, about 4:1, about 3:1, about 2:1, about 1:1, about 1:2, about 1:3, about 1:4, about 1:5, about 1:6, about 1:7, about 1:8, about 1:9, about 1:10, about 1:15, about 1:20, or about 1:25; - an M69 to N136 modification, substitution, and / or replacement ratio of about 25:1, about 20:1, about 15:1, about 10:1, about 9:1, about 8:1, about 7:1, about 6:1, about 5:1, about 4:1, about 3:1, about 2:1, about 1:1, about 1:2, about 1:3, about 1:4, about 1:5, about 1:6, about 1:7, about 1:8, about 1:9, about 1:10, about 1:15, about 1:20, or about 1:25; - an M69 to N138 modification, substitution, and / or replacement ratio of about 25:1, about 20:1, about 15:1, about 10:1, about 9:1, about 8:1, about 7:1, about 6:1, about 5:1, about 4:1, about 3:1, about 2:1, about 1:1, about 1:2, about 1:3, about 1:4, about 1:5, about 1:6, about 1:7, about 1:8, about 1:9, about 1:10, about 1:15, about 1:20, or about 1:25; a ratio of M69 to Q149 and modifications, substitutions, and / or substitutions of about 25:1, about 20:1, about 15:1, about 10:1, about 9:1, about 8:1, about 7:1, about 6:1, about 5:1, about 4:1, about 3:1, about 2:1, about 1:1, about 1:2, about 1:3, about 1:4, about 1:5, about 1:6, about 1:7, about 1:8, about 1:9, about 1:10, about 1:15, about 1:20, or about 1:25; - an M69 to N186 modification, substitution, and / or replacement ratio of about 25:1, about 20:1, about 15:1, about 10:1, about 9:1, about 8:1, about 7:1, about 6:1, about 5:1, about 4:1, about 3:1, about 2:1, about 1:1, about 1:2, about 1:3, about 1:4, about 1:5, about 1:6, about 1:7, about 1:8, about 1:9, about 1:10, about 1:15, about 1:20, or about 1:25; - an M69 to N200 modification, substitution, and / or replacement ratio of about 25:1, about 20:1, about 15:1, about 10:1, about 9:1, about 8:1, about 7:1, about 6:1, about 5:1, about 4:1, about 3:1, about 2:1, about 1:1, about 1:2, about 1:3, about 1:4, about 1:5, about 1:6, about 1:7, about 1:8, about 1:9, about 1:10, about 1:15, about 1:20, or about 1:25; a ratio of M69 to Q202 modification, substitution, and / or replacement of about 25:1, about 20:1, about 15:1, about 10:1, about 9:1, about 8:1, about 7:1, about 6:1, about 5:1, about 4:1, about 3:1, about 2:1, about 1:1, about 1:2, about 1:3, about 1:4, about 1:5, about 1:6, about 1:7, about 1:8, about 1:9, about 1:10, about 1:15, about 1:20, or about 1:25; - a ratio of M69 to N204 modification, substitution, and / or replacement of about 25:1, about 20:1, about 15:1, about 10:1, about 9:1, about 8:1, about 7:1, about 6:1, about 5:1, about 4:1, about 3:1, about 2:1, about 1:1, about 1:2, about 1:3, about 1:4, about 1:5, about 1:6, about 1:7, about 1:8, about 1:9, about 1:10, about 1:15, about 1:20, or about 1:25; - a ratio of M69 to N240 modification, substitution, and / or replacement of about 25:1, about 20:1, about 15:1, about 10:1, about 9:1, about 8:1, about 7:1, about 6:1, about 5:1, about 4:1, about 3:1, about 2:1, about 1:1, about 1:2, about 1:3, about 1:4, about 1:5, about 1:6, about 1:7, about 1:8, about 1:9, about 1:10, about 1:15, about 1:20, or about 1:25; - an M69 to N248 modification, substitution, and / or replacement ratio of about 25:1, about 20:1, about 15:1, about 10:1, about 9:1, about 8:1, about 7:1, about 6:1, about 5:1, about 4:1, about 3:1, about 2:1, about 1:1, about 1:2, about 1:3, about 1:4, about 1:5, about 1:6, about 1:7, about 1:8, about 1:9, about 1:10, about 1:15, about 1:20, or about 1:25; - a ratio of M69 to Q255 modification, substitution, and / or replacement of about 25:1, about 20:1, about 15:1, about 10:1, about 9:1, about 8:1, about 7:1, about 6:1, about 5:1, about 4:1, about 3:1, about 2:1, about 1:1, about 1:2, about 1:3, about 1:4, about 1:5, about 1:6, about 1:7, about 1:8, about 1:9, about 1:10, about 1:15, about 1:20, or about 1:25; a N105 to N108 modification, substitution, and / or replacement ratio of about 25:1, about 20:1, about 15:1, about 10:1, about 9:1, about 8:1, about 7:1, about 6:1, about 5:1, about 4:1, about 3:1, about 2:1, about 1:1, about 1:2, about 1:3, about 1:4, about 1:5, about 1:6, about 1:7, about 1:8, about 1:9, about 1:10, about 1:15, about 1:20, or about 1:25; a N105 to N118 modification, substitution, and / or replacement ratio of about 25:1, about 20:1, about 15:1, about 10:1, about 9:1, about 8:1, about 7:1, about 6:1, about 5:1, about 4:1, about 3:1, about 2:1, about 1:1, about 1:2, about 1:3, about 1:4, about 1:5, about 1:6, about 1:7, about 1:8, about 1:9, about 1:10, about 1:15, about 1:20, or about 1:25; a N105 to N136 modification, substitution, and / or replacement ratio of about 25:1, about 20:1, about 15:1, about 10:1, about 9:1, about 8:1, about 7:1, about 6:1, about 5:1, about 4:1, about 3:1, about 2:1, about 1:1, about 1:2, about 1:3, about 1:4, about 1:5, about 1:6, about 1:7, about 1:8, about 1:9, about 1:10, about 1:15, about 1:20, or about 1:25; a N105 to N138 modification, substitution, and / or replacement ratio of about 25:1, about 20:1, about 15:1, about 10:1, about 9:1, about 8:1, about 7:1, about 6:1, about 5:1, about 4:1, about 3:1, about 2:1, about 1:1, about 1:2, about 1:3, about 1:4, about 1:5, about 1:6, about 1:7, about 1:8, about 1:9, about 1:10, about 1:15, about 1:20, or about 1:25; a N105 to Q149 modification, substitution, and / or replacement ratio of about 25:1, about 20:1, about 15:1, about 10:1, about 9:1, about 8:1, about 7:1, about 6:1, about 5:1, about 4:1, about 3:1, about 2:1, about 1:1, about 1:2, about 1:3, about 1:4, about 1:5, about 1:6, about 1:7, about 1:8, about 1:9, about 1:10, about 1:15, about 1:20, or about 1:25; a N105 to N186 modification, substitution, and / or replacement ratio of about 25:1, about 20:1, about 15:1, about 10:1, about 9:1, about 8:1, about 7:1, about 6:1, about 5:1, about 4:1, about 3:1, about 2:1, about 1:1, about 1:2, about 1:3, about 1:4, about 1:5, about 1:6, about 1:7, about 1:8, about 1:9, about 1:10, about 1:15, about 1:20, or about 1:25; - a N105 to N200 modification, substitution, and / or replacement ratio of about 25:1, about 20:1, about 15:1, about 10:1, about 9:1, about 8:1, about 7:1, about 6:1, about 5:1, about 4:1, about 3:1, about 2:1, about 1:1, about 1:2, about 1:3, about 1:4, about 1:5, about 1:6, about 1:7, about 1:8, about 1:9, about 1:10, about 1:15, about 1:20, or about 1:25; a modification, substitution, and / or replacement ratio of Q202 to N105 of about 25:1, about 20:1, about 15:1, about 10:1, about 9:1, about 8:1, about 7:1, about 6:1, about 5:1, about 4:1, about 3:1, about 2:1, about 1:1, about 1:2, about 1:3, about 1:4, about 1:5, about 1:6, about 1:7, about 1:8, about 1:9, about 1:10, about 1:15, about 1:20, or about 1:25; a N105 to N204 modification, substitution, and / or replacement ratio of about 25:1, about 20:1, about 15:1, about 10:1, about 9:1, about 8:1, about 7:1, about 6:1, about 5:1, about 4:1, about 3:1, about 2:1, about 1:1, about 1:2, about 1:3, about 1:4, about 1:5, about 1:6, about 1:7, about 1:8, about 1:9, about 1:10, about 1:15, about 1:20, or about 1:25; - a N105 to N240 modification, substitution, and / or replacement ratio of about 25:1, about 20:1, about 15:1, about 10:1, about 9:1, about 8:1, about 7:1, about 6:1, about 5:1, about 4:1, about 3:1, about 2:1, about 1:1, about 1:2, about 1:3, about 1:4, about 1:5, about 1:6, about 1:7, about 1:8, about 1:9, about 1:10, about 1:15, about 1:20, or about 1:25; a N105 to N248 modification, substitution, and / or replacement ratio of about 25:1, about 20:1, about 15:1, about 10:1, about 9:1, about 8:1, about 7:1, about 6:1, about 5:1, about 4:1, about 3:1, about 2:1, about 1:1, about 1:2, about 1:3, about 1:4, about 1:5, about 1:6, about 1:7, about 1:8, about 1:9, about 1:10, about 1:15, about 1:20, or about 1:25; a N105 to Q255 modification, substitution, and / or replacement ratio of about 25:1, about 20:1, about 15:1, about 10:1, about 9:1, about 8:1, about 7:1, about 6:1, about 5:1, about 4:1, about 3:1, about 2:1, about 1:1, about 1:2, about 1:3, about 1:4, about 1:5, about 1:6, about 1:7, about 1:8, about 1:9, about 1:10, about 1:15, about 1:20, or about 1:25; a N108 to N118 modification, substitution, and / or replacement ratio of about 25:1, about 20:1, about 15:1, about 10:1, about 9:1, about 8:1, about 7:1, about 6:1, about 5:1, about 4:1, about 3:1, about 2:1, about 1:1, about 1:2, about 1:3, about 1:4, about 1:5, about 1:6, about 1:7, about 1:8, about 1:9, about 1:10, about 1:15, about 1:20, or about 1:25; a N108 to N136 modification, substitution, and / or replacement ratio of about 25:1, about 20:1, about 15:1, about 10:1, about 9:1, about 8:1, about 7:1, about 6:1, about 5:1, about 4:1, about 3:1, about 2:1, about 1:1, about 1:2, about 1:3, about 1:4, about 1:5, about 1:6, about 1:7, about 1:8, about 1:9, about 1:10, about 1:15, about 1:20, or about 1:25; a N108 to N138 modification, substitution, and / or replacement ratio of about 25:1, about 20:1, about 15:1, about 10:1, about 9:1, about 8:1, about 7:1, about 6:1, about 5:1, about 4:1, about 3:1, about 2:1, about 1:1, about 1:2, about 1:3, about 1:4, about 1:5, about 1:6, about 1:7, about 1:8, about 1:9, about 1:10, about 1:15, about 1:20, or about 1:25; a N108 to Q149 modification, substitution, and / or replacement ratio of about 25:1, about 20:1, about 15:1, about 10:1, about 9:1, about 8:1, about 7:1, about 6:1, about 5:1, about 4:1, about 3:1, about 2:1, about 1:1, about 1:2, about 1:3, about 1:4, about 1:5, about 1:6, about 1:7, about 1:8, about 1:9, about 1:10, about 1:15, about 1:20, or about 1:25; - a N108 to N186 modification, substitution, and / or replacement ratio of about 25:1, about 20:1, about 15:1, about 10:1, about 9:1, about 8:1, about 7:1, about 6:1, about 5:1, about 4:1, about 3:1, about 2:1, about 1:1, about 1:2, about 1:3, about 1:4, about 1:5, about 1:6, about 1:7, about 1:8, about 1:9, about 1:10, about 1:15, about 1:20, or about 1:25; - a N108 to N200 modification, substitution, and / or substitution ratio of about 25:1, about 20:1, about 15:1, about 10:1, about 9:1, about 8:1, about 7:1, about 6:1, about 5:1, about 4:1, about 3:1, about 2:1, about 1:1, about 1:2, about 1:3, about 1:4, about 1:5, about 1:6, about 1:7, about 1:8, about 1:9, about 1:10, about 1:15, about 1:20, or about 1:25; a N108 to Q202 modification, substitution, and / or substitution ratio of about 25:1, about 20:1, about 15:1, about 10:1, about 9:1, about 8:1, about 7:1, about 6:1, about 5:1, about 4:1, about 3:1, about 2:1, about 1:1, about 1:2, about 1:3, about 1:4, about 1:5, about 1:6, about 1:7, about 1:8, about 1:9, about 1:10, about 1:15, about 1:20, or about 1:25; a N108 to N204 modification, substitution, and / or replacement ratio of about 25:1, about 20:1, about 15:1, about 10:1, about 9:1, about 8:1, about 7:1, about 6:1, about 5:1, about 4:1, about 3:1, about 2:1, about 1:1, about 1:2, about 1:3, about 1:4, about 1:5, about 1:6, about 1:7, about 1:8, about 1:9, about 1:10, about 1:15, about 1:20, or about 1:25; a N108 to N240 modification, substitution, and / or replacement ratio of about 25:1, about 20:1, about 15:1, about 10:1, about 9:1, about 8:1, about 7:1, about 6:1, about 5:1, about 4:1, about 3:1, about 2:1, about 1:1, about 1:2, about 1:3, about 1:4, about 1:5, about 1:6, about 1:7, about 1:8, about 1:9, about 1:10, about 1:15, about 1:20, or about 1:25; a N108 to N248 modification, substitution, and / or replacement ratio of about 25:1, about 20:1, about 15:1, about 10:1, about 9:1, about 8:1, about 7:1, about 6:1, about 5:1, about 4:1, about 3:1, about 2:1, about 1:1, about 1:2, about 1:3, about 1:4, about 1:5, about 1:6, about 1:7, about 1:8, about 1:9, about 1:10, about 1:15, about 1:20, or about 1:25; a N108 to Q255 modification, substitution, and / or substitution ratio of about 25:1, about 20:1, about 15:1, about 10:1, about 9:1, about 8:1, about 7:1, about 6:1, about 5:1, about 4:1, about 3:1, about 2:1, about 1:1, about 1:2, about 1:3, about 1:4, about 1:5, about 1:6, about 1:7, about 1:8, about 1:9, about 1:10, about 1:15, about 1:20, or about 1:25; a N118 to N136 modification, substitution, and / or replacement ratio of about 25:1, about 20:1, about 15:1, about 10:1, about 9:1, about 8:1, about 7:1, about 6:1, about 5:1, about 4:1, about 3:1, about 2:1, about 1:1, about 1:2, about 1:3, about 1:4, about 1:5, about 1:6, about 1:7, about 1:8, about 1:9, about 1:10, about 1:15, about 1:20, or about 1:25; a N118 to N138 modification, substitution, and / or replacement ratio of about 25:1, about 20:1, about 15:1, about 10:1, about 9:1, about 8:1, about 7:1, about 6:1, about 5:1, about 4:1, about 3:1, about 2:1, about 1:1, about 1:2, about 1:3, about 1:4, about 1:5, about 1:6, about 1:7, about 1:8, about 1:9, about 1:10, about 1:15, about 1:20, or about 1:25; a N118 to Q149 modification, substitution, and / or replacement ratio of about 25:1, about 20:1, about 15:1, about 10:1, about 9:1, about 8:1, about 7:1, about 6:1, about 5:1, about 4:1, about 3:1, about 2:1, about 1:1, about 1:2, about 1:3, about 1:4, about 1:5, about 1:6, about 1:7, about 1:8, about 1:9, about 1:10, about 1:15, about 1:20, or about 1:25; a N118 to N186 modification, substitution, and / or replacement ratio of about 25:1, about 20:1, about 15:1, about 10:1, about 9:1, about 8:1, about 7:1, about 6:1, about 5:1, about 4:1, about 3:1, about 2:1, about 1:1, about 1:2, about 1:3, about 1:4, about 1:5, about 1:6, about 1:7, about 1:8, about 1:9, about 1:10, about 1:15, about 1:20, or about 1:25; - a N118 to N200 modification, substitution, and / or replacement ratio of about 25:1, about 20:1, about 15:1, about 10:1, about 9:1, about 8:1, about 7:1, about 6:1, about 5:1, about 4:1, about 3:1, about 2:1, about 1:1, about 1:2, about 1:3, about 1:4, about 1:5, about 1:6, about 1:7, about 1:8, about 1:9, about 1:10, about 1:15, about 1:20, or about 1:25; a N118 to Q202 modification, substitution, and / or substitution ratio of about 25:1, about 20:1, about 15:1, about 10:1, about 9:1, about 8:1, about 7:1, about 6:1, about 5:1, about 4:1, about 3:1, about 2:1, about 1:1, about 1:2, about 1:3, about 1:4, about 1:5, about 1:6, about 1:7, about 1:8, about 1:9, about 1:10, about 1:15, about 1:20, or about 1:25; a N118 to N204 modification, substitution, and / or replacement ratio of about 25:1, about 20:1, about 15:1, about 10:1, about 9:1, about 8:1, about 7:1, about 6:1, about 5:1, about 4:1, about 3:1, about 2:1, about 1:1, about 1:2, about 1:3, about 1:4, about 1:5, about 1:6, about 1:7, about 1:8, about 1:9, about 1:10, about 1:15, about 1:20, or about 1:25; a N118 to N240 modification, substitution, and / or replacement ratio of about 25:1, about 20:1, about 15:1, about 10:1, about 9:1, about 8:1, about 7:1, about 6:1, about 5:1, about 4:1, about 3:1, about 2:1, about 1:1, about 1:2, about 1:3, about 1:4, about 1:5, about 1:6, about 1:7, about 1:8, about 1:9, about 1:10, about 1:15, about 1:20, or about 1:25; a N118 to N248 modification, substitution, and / or replacement ratio of about 25:1, about 20:1, about 15:1, about 10:1, about 9:1, about 8:1, about 7:1, about 6:1, about 5:1, about 4:1, about 3:1, about 2:1, about 1:1, about 1:2, about 1:3, about 1:4, about 1:5, about 1:6, about 1:7, about 1:8, about 1:9, about 1:10, about 1:15, about 1:20, or about 1:25; a N118 to Q255 modification, substitution, and / or replacement ratio of about 25:1, about 20:1, about 15:1, about 10:1, about 9:1, about 8:1, about 7:1, about 6:1, about 5:1, about 4:1, about 3:1, about 2:1, about 1:1, about 1:2, about 1:3, about 1:4, about 1:5, about 1:6, about 1:7, about 1:8, about 1:9, about 1:10, about 1:15, about 1:20, or about 1:25; a N136 to N138 modification, substitution, and / or replacement ratio of about 25:1, about 20:1, about 15:1, about 10:1, about 9:1, about 8:1, about 7:1, about 6:1, about 5:1, about 4:1, about 3:1, about 2:1, about 1:1, about 1:2, about 1:3, about 1:4, about 1:5, about 1:6, about 1:7, about 1:8, about 1:9, about 1:10, about 1:15, about 1:20, or about 1:25; - a N136 to Q149 modification, substitution, and / or replacement ratio of about 25:1, about 20:1, about 15:1, about 10:1, about 9:1, about 8:1, about 7:1, about 6:1, about 5:1, about 4:1, about 3:1, about 2:1, about 1:1, about 1:2, about 1:3, about 1:4, about 1:5, about 1:6, about 1:7, about 1:8, about 1:9, about 1:10, about 1:15, about 1:20, or about 1:25; - a N136 to N186 modification, substitution, and / or replacement ratio of about 25:1, about 20:1, about 15:1, about 10:1, about 9:1, about 8:1, about 7:1, about 6:1, about 5:1, about 4:1, about 3:1, about 2:1, about 1:1, about 1:2, about 1:3, about 1:4, about 1:5, about 1:6, about 1:7, about 1:8, about 1:9, about 1:10, about 1:15, about 1:20, or about 1:25; - a N136 to N200 modification, substitution, and / or replacement ratio of about 25:1, about 20:1, about 15:1, about 10:1, about 9:1, about 8:1, about 7:1, about 6:1, about 5:1, about 4:1, about 3:1, about 2:1, about 1:1, about 1:2, about 1:3, about 1:4, about 1:5, about 1:6, about 1:7, about 1:8, about 1:9, about 1:10, about 1:15, about 1:20, or about 1:25; a N136 to Q202 modification, substitution, and / or substitution ratio of about 25:1, about 20:1, about 15:1, about 10:1, about 9:1, about 8:1, about 7:1, about 6:1, about 5:1, about 4:1, about 3:1, about 2:1, about 1:1, about 1:2, about 1:3, about 1:4, about 1:5, about 1:6, about 1:7, about 1:8, about 1:9, about 1:10, about 1:15, about 1:20, or about 1:25; a N136 to N204 modification, substitution, and / or replacement ratio of about 25:1, about 20:1, about 15:1, about 10:1, about 9:1, about 8:1, about 7:1, about 6:1, about 5:1, about 4:1, about 3:1, about 2:1, about 1:1, about 1:2, about 1:3, about 1:4, about 1:5, about 1:6, about 1:7, about 1:8, about 1:9, about 1:10, about 1:15, about 1:20, or about 1:25; an N136 to N240 modification, substitution, and / or replacement ratio of about 25:1, about 20:1, about 15:1, about 10:1, about 9:1, about 8:1, about 7:1, about 6:1, about 5:1, about 4:1, about 3:1, about 2:1, about 1:1, about 1:2, about 1:3, about 1:4, about 1:5, about 1:6, about 1:7, about 1:8, about 1:9, about 1:10, about 1:15, about 1:20, or about 1:25; - a N136 to N248 modification, substitution, and / or replacement ratio of about 25:1, about 20:1, about 15:1, about 10:1, about 9:1, about 8:1, about 7:1, about 6:1, about 5:1, about 4:1, about 3:1, about 2:1, about 1:1, about 1:2, about 1:3, about 1:4, about 1:5, about 1:6, about 1:7, about 1:8, about 1:9, about 1:10, about 1:15, about 1:20, or about 1:25; a N136 to Q255 modification, substitution, and / or replacement ratio of about 25:1, about 20:1, about 15:1, about 10:1, about 9:1, about 8:1, about 7:1, about 6:1, about 5:1, about 4:1, about 3:1, about 2:1, about 1:1, about 1:2, about 1:3, about 1:4, about 1:5, about 1:6, about 1:7, about 1:8, about 1:9, about 1:10, about 1:15, about 1:20, or about 1:25; a N138 to Q149 modification, substitution, and / or replacement ratio of about 25:1, about 20:1, about 15:1, about 10:1, about 9:1, about 8:1, about 7:1, about 6:1, about 5:1, about 4:1, about 3:1, about 2:1, about 1:1, about 1:2, about 1:3, about 1:4, about 1:5, about 1:6, about 1:7, about 1:8, about 1:9, about 1:10, about 1:15, about 1:20, or about 1:25; - a N138 to N186 modification, substitution, and / or replacement ratio of about 25:1, about 20:1, about 15:1, about 10:1, about 9:1, about 8:1, about 7:1, about 6:1, about 5:1, about 4:1, about 3:1, about 2:1, about 1:1, about 1:2, about 1:3, about 1:4, about 1:5, about 1:6, about 1:7, about 1:8, about 1:9, about 1:10, about 1:15, about 1:20, or about 1:25; - a N138 to N200 modification, substitution, and / or replacement ratio of about 25:1, about 20:1, about 15:1, about 10:1, about 9:1, about 8:1, about 7:1, about 6:1, about 5:1, about 4:1, about 3:1, about 2:1, about 1:1, about 1:2, about 1:3, about 1:4, about 1:5, about 1:6, about 1:7, about 1:8, about 1:9, about 1:10, about 1:15, about 1:20, or about 1:25; a N138 to Q202 modification, substitution, and / or replacement ratio of about 25:1, about 20:1, about 15:1, about 10:1, about 9:1, about 8:1, about 7:1, about 6:1, about 5:1, about 4:1, about 3:1, about 2:1, about 1:1, about 1:2, about 1:3, about 1:4, about 1:5, about 1:6, about 1:7, about 1:8, about 1:9, about 1:10, about 1:15, about 1:20, or about 1:25; a N138 to N204 modification, substitution, and / or replacement ratio of about 25:1, about 20:1, about 15:1, about 10:1, about 9:1, about 8:1, about 7:1, about 6:1, about 5:1, about 4:1, about 3:1, about 2:1, about 1:1, about 1:2, about 1:3, about 1:4, about 1:5, about 1:6, about 1:7, about 1:8, about 1:9, about 1:10, about 1:15, about 1:20, or about 1:25; an N138 to N240 modification, substitution, and / or replacement ratio of about 25:1, about 20:1, about 15:1, about 10:1, about 9:1, about 8:1, about 7:1, about 6:1, about 5:1, about 4:1, about 3:1, about 2:1, about 1:1, about 1:2, about 1:3, about 1:4, about 1:5, about 1:6, about 1:7, about 1:8, about 1:9, about 1:10, about 1:15, about 1:20, or about 1:25; a N138 to N248 modification, substitution, and / or replacement ratio of about 25:1, about 20:1, about 15:1, about 10:1, about 9:1, about 8:1, about 7:1, about 6:1, about 5:1, about 4:1, about 3:1, about 2:1, about 1:1, about 1:2, about 1:3, about 1:4, about 1:5, about 1:6, about 1:7, about 1:8, about 1:9, about 1:10, about 1:15, about 1:20, or about 1:25; a N138 to Q255 modification, substitution, and / or replacement ratio of about 25:1, about 20:1, about 15:1, about 10:1, about 9:1, about 8:1, about 7:1, about 6:1, about 5:1, about 4:1, about 3:1, about 2:1, about 1:1, about 1:2, about 1:3, about 1:4, about 1:5, about 1:6, about 1:7, about 1:8, about 1:9, about 1:10, about 1:15, about 1:20, or about 1:25; a Q149 to N186 modification, substitution, and / or replacement ratio of about 25:1, about 20:1, about 15:1, about 10:1, about 9:1, about 8:1, about 7:1, about 6:1, about 5:1, about 4:1, about 3:1, about 2:1, about 1:1, about 1:2, about 1:3, about 1:4, about 1:5, about 1:6, about 1:7, about 1:8, about 1:9, about 1:10, about 1:15, about 1:20, or about 1:25; a Q149 to N200 modification, substitution, and / or replacement ratio of about 25:1, about 20:1, about 15:1, about 10:1, about 9:1, about 8:1, about 7:1, about 6:1, about 5:1, about 4:1, about 3:1, about 2:1, about 1:1, about 1:2, about 1:3, about 1:4, about 1:5, about 1:6, about 1:7, about 1:8, about 1:9, about 1:10, about 1:15, about 1:20, or about 1:25; a modification, substitution, and / or replacement ratio of Q149 to Q202 of about 25:1, about 20:1, about 15:1, about 10:1, about 9:1, about 8:1, about 7:1, about 6:1, about 5:1, about 4:1, about 3:1, about 2:1, about 1:1, about 1:2, about 1:3, about 1:4, about 1:5, about 1:6, about 1:7, about 1:8, about 1:9, about 1:10, about 1:15, about 1:20, or about 1:25; a Q149 to N204 modification, substitution, and / or replacement ratio of about 25:1, about 20:1, about 15:1, about 10:1, about 9:1, about 8:1, about 7:1, about 6:1, about 5:1, about 4:1, about 3:1, about 2:1, about 1:1, about 1:2, about 1:3, about 1:4, about 1:5, about 1:6, about 1:7, about 1:8, about 1:9, about 1:10, about 1:15, about 1:20, or about 1:25; a Q149 to N240 modification, substitution, and / or replacement ratio of about 25:1, about 20:1, about 15:1, about 10:1, about 9:1, about 8:1, about 7:1, about 6:1, about 5:1, about 4:1, about 3:1, about 2:1, about 1:1, about 1:2, about 1:3, about 1:4, about 1:5, about 1:6, about 1:7, about 1:8, about 1:9, about 1:10, about 1:15, about 1:20, or about 1:25; a Q149 to N248 modification, substitution, and / or replacement ratio of about 25:1, about 20:1, about 15:1, about 10:1, about 9:1, about 8:1, about 7:1, about 6:1, about 5:1, about 4:1, about 3:1, about 2:1, about 1:1, about 1:2, about 1:3, about 1:4, about 1:5, about 1:6, about 1:7, about 1:8, about 1:9, about 1:10, about 1:15, about 1:20, or about 1:25; a Q149 to Q255 modification, substitution, and / or replacement ratio of about 25:1, about 20:1, about 15:1, about 10:1, about 9:1, about 8:1, about 7:1, about 6:1, about 5:1, about 4:1, about 3:1, about 2:1, about 1:1, about 1:2, about 1:3, about 1:4, about 1:5, about 1:6, about 1:7, about 1:8, about 1:9, about 1:10, about 1:15, about 1:20, or about 1:25; - a N186 to N200 modification, substitution, and / or replacement ratio of about 25:1, about 20:1, about 15:1, about 10:1, about 9:1, about 8:1, about 7:1, about 6:1, about 5:1, about 4:1, about 3:1, about 2:1, about 1:1, about 1:2, about 1:3, about 1:4, about 1:5, about 1:6, about 1:7, about 1:8, about 1:9, about 1:10, about 1:15, about 1:20, or about 1:25; a N186 to Q202 modification, substitution, and / or replacement ratio of about 25:1, about 20:1, about 15:1, about 10:1, about 9:1, about 8:1, about 7:1, about 6:1, about 5:1, about 4:1, about 3:1, about 2:1, about 1:1, about 1:2, about 1:3, about 1:4, about 1:5, about 1:6, about 1:7, about 1:8, about 1:9, about 1:10, about 1:15, about 1:20, or about 1:25; - a N186 to N204 modification, substitution, and / or replacement ratio of about 25:1, about 20:1, about 15:1, about 10:1, about 9:1, about 8:1, about 7:1, about 6:1, about 5:1, about 4:1, about 3:1, about 2:1, about 1:1, about 1:2, about 1:3, about 1:4, about 1:5, about 1:6, about 1:7, about 1:8, about 1:9, about 1:10, about 1:15, about 1:20, or about 1:25; - a N186 to N240 modification, substitution, and / or replacement ratio of about 25:1, about 20:1, about 15:1, about 10:1, about 9:1, about 8:1, about 7:1, about 6:1, about 5:1, about 4:1, about 3:1, about 2:1, about 1:1, about 1:2, about 1:3, about 1:4, about 1:5, about 1:6, about 1:7, about 1:8, about 1:9, about 1:10, about 1:15, about 1:20, or about 1:25; - a N186 to N248 modification, substitution, and / or replacement ratio of about 25:1, about 20:1, about 15:1, about 10:1, about 9:1, about 8:1, about 7:1, about 6:1, about 5:1, about 4:1, about 3:1, about 2:1, about 1:1, about 1:2, about 1:3, about 1:4, about 1:5, about 1:6, about 1:7, about 1:8, about 1:9, about 1:10, about 1:15, about 1:20, or about 1:25; a N186 to Q255 modification, substitution, and / or replacement ratio of about 25:1, about 20:1, about 15:1, about 10:1, about 9:1, about 8:1, about 7:1, about 6:1, about 5:1, about 4:1, about 3:1, about 2:1, about 1:1, about 1:2, about 1:3, about 1:4, about 1:5, about 1:6, about 1:7, about 1:8, about 1:9, about 1:10, about 1:15, about 1:20, or about 1:25; a N200 to Q202 modification, substitution, and / or replacement ratio of about 25:1, about 20:1, about 15:1, about 10:1, about 9:1, about 8:1, about 7:1, about 6:1, about 5:1, about 4:1, about 3:1, about 2:1, about 1:1, about 1:2, about 1:3, about 1:4, about 1:5, about 1:6, about 1:7, about 1:8, about 1:9, about 1:10, about 1:15, about 1:20, or about 1:25; a modification, substitution, and / or replacement ratio of N200 to N204 of about 25:1, about 20:1, about 15:1, about 10:1, about 9:1, about 8:1, about 7:1, about 6:1, about 5:1, about 4:1, about 3:1, about 2:1, about 1:1, about 1:2, about 1:3, about 1:4, about 1:5, about 1:6, about 1:7, about 1:8, about 1:9, about 1:10, about 1:15, about 1:20, or about 1:25; a modification, substitution, and / or replacement ratio of N200 to N240 of about 25:1, about 20:1, about 15:1, about 10:1, about 9:1, about 8:1, about 7:1, about 6:1, about 5:1, about 4:1, about 3:1, about 2:1, about 1:1, about 1:2, about 1:3, about 1:4, about 1:5, about 1:6, about 1:7, about 1:8, about 1:9, about 1:10, about 1:15, about 1:20, or about 1:25; - a modification, substitution, and / or replacement ratio of N200 to N248 of about 25:1, about 20:1, about 15:1, about 10:1, about 9:1, about 8:1, about 7:1, about 6:1, about 5:1, about 4:1, about 3:1, about 2:1, about 1:1, about 1:2, about 1:3, about 1:4, about 1:5, about 1:6, about 1:7, about 1:8, about 1:9, about 1:10, about 1:15, about 1:20, or about 1:25; - a N200 to Q255 modification, substitution, and / or replacement ratio of about 25:1, about 20:1, about 15:1, about 10:1, about 9:1, about 8:1, about 7:1, about 6:1, about 5:1, about 4:1, about 3:1, about 2:1, about 1:1, about 1:2, about 1:3, about 1:4, about 1:5, about 1:6, about 1:7, about 1:8, about 1:9, about 1:10, about 1:15, about 1:20, or about 1:25; a Q202 to N204 modification, substitution, and / or replacement ratio of about 25:1, about 20:1, about 15:1, about 10:1, about 9:1, about 8:1, about 7:1, about 6:1, about 5:1, about 4:1, about 3:1, about 2:1, about 1:1, about 1:2, about 1:3, about 1:4, about 1:5, about 1:6, about 1:7, about 1:8, about 1:9, about 1:10, about 1:15, about 1:20, or about 1:25; a Q202 to N240 modification, substitution, and / or replacement ratio of about 25:1, about 20:1, about 15:1, about 10:1, about 9:1, about 8:1, about 7:1, about 6:1, about 5:1, about 4:1, about 3:1, about 2:1, about 1:1, about 1:2, about 1:3, about 1:4, about 1:5, about 1:6, about 1:7, about 1:8, about 1:9, about 1:10, about 1:15, about 1:20, or about 1:25; a modification, substitution, and / or replacement ratio of Q202 to N248 of about 25:1, about 20:1, about 15:1, about 10:1, about 9:1, about 8:1, about 7:1, about 6:1, about 5:1, about 4:1, about 3:1, about 2:1, about 1:1, about 1:2, about 1:3, about 1:4, about 1:5, about 1:6, about 1:7, about 1:8, about 1:9, about 1:10, about 1:15, about 1:20, or about 1:25; a modification, substitution, and / or substitution ratio of Q202 to Q255 of about 25:1, about 20:1, about 15:1, about 10:1, about 9:1, about 8:1, about 7:1, about 6:1, about 5:1, about 4:1, about 3:1, about 2:1, about 1:1, about 1:2, about 1:3, about 1:4, about 1:5, about 1:6, about 1:7, about 1:8, about 1:9, about 1:10, about 1:15, about 1:20, or about 1:25; a N204 to N240 modification, substitution, and / or replacement ratio of about 25:1, about 20:1, about 15:1, about 10:1, about 9:1, about 8:1, about 7:1, about 6:1, about 5:1, about 4:1, about 3:1, about 2:1, about 1:1, about 1:2, about 1:3, about 1:4, about 1:5, about 1:6, about 1:7, about 1:8, about 1:9, about 1:10, about 1:15, about 1:20, or about 1:25; a N204 to N248 modification, substitution, and / or replacement ratio of about 25:1, about 20:1, about 15:1, about 10:1, about 9:1, about 8:1, about 7:1, about 6:1, about 5:1, about 4:1, about 3:1, about 2:1, about 1:1, about 1:2, about 1:3, about 1:4, about 1:5, about 1:6, about 1:7, about 1:8, about 1:9, about 1:10, about 1:15, about 1:20, or about 1:25; a N204 to Q255 modification, substitution, and / or replacement ratio of about 25:1, about 20:1, about 15:1, about 10:1, about 9:1, about 8:1, about 7:1, about 6:1, about 5:1, about 4:1, about 3:1, about 2:1, about 1:1, about 1:2, about 1:3, about 1:4, about 1:5, about 1:6, about 1:7, about 1:8, about 1:9, about 1:10, about 1:15, about 1:20, or about 1:25; - a N240 to N248 modification, substitution, and / or replacement ratio of about 25:1, about 20:1, about 15:1, about 10:1, about 9:1, about 8:1, about 7:1, about 6:1, about 5:1, about 4:1, about 3:1, about 2:1, about 1:1, about 1:2, about 1:3, about 1:4, about 1:5, about 1:6, about 1:7, about 1:8, about 1:9, about 1:10, about 1:15, about 1:20, or about 1:25; an N240 to Q255 modification, substitution, and / or replacement ratio of about 25:1, about 20:1, about 15:1, about 10:1, about 9:1, about 8:1, about 7:1, about 6:1, about 5:1, about 4:1, about 3:1, about 2:1, about 1:1, about 1:2, about 1:3, about 1:4, about 1:5, about 1:6, about 1:7, about 1:8, about 1:9, about 1:10, about 1:15, about 1:20, or about 1:25; a N248 to Q255 modification, substitution, and / or replacement ratio of about 25:1, about 20:1, about 15:1, about 10:1, about 9:1, about 8:1, about 7:1, about 6:1, about 5:1, about 4:1, about 3:1, about 2:1, about 1:1, about 1:2, about 1:3, about 1:4, about 1:5, about 1:6, about 1:7, about 1:8, about 1:9, about 1:10, about 1:15, about 1:20, or about 1:25;

[0012] In some embodiments, the present disclosure provides compositions that include a plurality of peptides or protein fragments, e.g., a plurality of fibroin light and heavy chain peptides or fibroin light and heavy chain fragments, and that include a plurality of amino acid modifications, substitutions, and / or substitutions, and further include, at specific positions, ratios of one or more modifications, substitutions, and / or substitutions selected from the following: - a ratio of modifications, substitutions, and / or substitutions of light chain N23 to heavy chain Q58, M64, N68, N70, N77, M80, N93, M103, Q125, N132, Q139, Q275, N4191, Q5216, or N5262 of about 25:1, about 20:1, about 15:1, about 10:1, about 9:1, about 8:1, about 7:1, about 6:1, about 5:1, about 4:1, about 3:1, about 2:1, about 1:1, about 1:2, about 1:3, about 1:4, about 1:5, about 1:6, about 1:7, about 1:8, about 1:9, about 1:10, about 1:15, about 1:20, or about 1:25; - a ratio of modifications, substitutions, and / or substitutions of light chain N24 to heavy chain Q58, M64, N68, N70, N77, M80, N93, M103, Q125, N132, Q139, Q275, N4191, Q5216, or N5262 of about 25:1, about 20:1, about 15:1, about 10:1, about 9:1, about 8:1, about 7:1, about 6:1, about 5:1, about 4:1, about 3:1, about 2:1, about 1:1, about 1:2, about 1:3, about 1:4, about 1:5, about 1:6, about 1:7, about 1:8, about 1:9, about 1:10, about 1:15, about 1:20, or about 1:25; - a ratio of modifications, substitutions, and / or substitutions of light chain N28 to heavy chain Q58, M64, N68, N70, N77, M80, N93, M103, Q125, N132, Q139, Q275, N4191, Q5216, or N5262 of about 25:1, about 20:1, about 15:1, about 10:1, about 9:1, about 8:1, about 7:1, about 6:1, about 5:1, about 4:1, about 3:1, about 2:1, about 1:1, about 1:2, about 1:3, about 1:4, about 1:5, about 1:6, about 1:7, about 1:8, about 1:9, about 1:10, about 1:15, about 1:20, or about 1:25; - a ratio of light chain M69 to heavy chain Q58, M64, N68, N70, N77, M80, N93, M103, Q125, N132, Q139, Q275, N4191, Q5216, or N5262 modifications, substitutions, and / or substitutions of about 25:1, about 20:1, about 15:1, about 10:1, about 9:1, about 8:1, about 7:1, about 6:1, about 5:1, about 4:1, about 3:1, about 2:1, about 1:1, about 1:2, about 1:3, about 1:4, about 1:5, about 1:6, about 1:7, about 1:8, about 1:9, about 1:10, about 1:15, about 1:20, or about 1:25; - a ratio of modifications, substitutions, and / or substitutions of light chain N105 to heavy chain Q58, M64, N68, N70, N77, M80, N93, M103, Q125, N132, Q139, Q275, N4191, Q5216, or N5262 of about 25:1, about 20:1, about 15:1, about 10:1, about 9:1, about 8:1, about 7:1, about 6:1, about 5:1, about 4:1, about 3:1, about 2:1, about 1:1, about 1:2, about 1:3, about 1:4, about 1:5, about 1:6, about 1:7, about 1:8, about 1:9, about 1:10, about 1:15, about 1:20, or about 1:25; - a ratio of modifications, substitutions, and / or substitutions of light chain N108 to heavy chain Q58, M64, N68, N70, N77, M80, N93, M103, Q125, N132, Q139, Q275, N4191, Q5216, or N5262 of about 25:1, about 20:1, about 15:1, about 10:1, about 9:1, about 8:1, about 7:1, about 6:1, about 5:1, about 4:1, about 3:1, about 2:1, about 1:1, about 1:2, about 1:3, about 1:4, about 1:5, about 1:6, about 1:7, about 1:8, about 1:9, about 1:10, about 1:15, about 1:20, or about 1:25; - a ratio of modifications, substitutions, and / or substitutions of light chain N118 to heavy chain Q58, M64, N68, N70, N77, M80, N93, M103, Q125, N132, Q139, Q275, N4191, Q5216, or N5262 of about 25:1, about 20:1, about 15:1, about 10:1, about 9:1, about 8:1, about 7:1, about 6:1, about 5:1, about 4:1, about 3:1, about 2:1, about 1:1, about 1:2, about 1:3, about 1:4, about 1:5, about 1:6, about 1:7, about 1:8, about 1:9, about 1:10, about 1:15, about 1:20, or about 1:25; - a ratio of modifications, substitutions, and / or substitutions of light chain N136 to heavy chain Q58, M64, N68, N70, N77, M80, N93, M103, Q125, N132, Q139, Q275, N4191, Q5216, or N5262 of about 25:1, about 20:1, about 15:1, about 10:1, about 9:1, about 8:1, about 7:1, about 6:1, about 5:1, about 4:1, about 3:1, about 2:1, about 1:1, about 1:2, about 1:3, about 1:4, about 1:5, about 1:6, about 1:7, about 1:8, about 1:9, about 1:10, about 1:15, about 1:20, or about 1:25; - a ratio of modifications, substitutions, and / or substitutions of light chain N138 to heavy chain Q58, M64, N68, N70, N77, M80, N93, M103, Q125, N132, Q139, Q275, N4191, Q5216, or N5262 of about 25:1, about 20:1, about 15:1, about 10:1, about 9:1, about 8:1, about 7:1, about 6:1, about 5:1, about 4:1, about 3:1, about 2:1, about 1:1, about 1:2, about 1:3, about 1:4, about 1:5, about 1:6, about 1:7, about 1:8, about 1:9, about 1:10, about 1:15, about 1:20, or about 1:25; - a ratio of modifications, substitutions, and / or substitutions of light chain Q149 to heavy chain Q58, M64, N68, N70, N77, M80, N93, M103, Q125, N132, Q139, Q275, N4191, Q5216, or N5262 of about 25:1, about 20:1, about 15:1, about 10:1, about 9:1, about 8:1, about 7:1, about 6:1, about 5:1, about 4:1, about 3:1, about 2:1, about 1:1, about 1:2, about 1:3, about 1:4, about 1:5, about 1:6, about 1:7, about 1:8, about 1:9, about 1:10, about 1:15, about 1:20, or about 1:25; - a ratio of modifications, substitutions, and / or substitutions of light chain N186 to heavy chain Q58, M64, N68, N70, N77, M80, N93, M103, Q125, N132, Q139, Q275, N4191, Q5216, or N5262 of about 25:1, about 20:1, about 15:1, about 10:1, about 9:1, about 8:1, about 7:1, about 6:1, about 5:1, about 4:1, about 3:1, about 2:1, about 1:1, about 1:2, about 1:3, about 1:4, about 1:5, about 1:6, about 1:7, about 1:8, about 1:9, about 1:10, about 1:15, about 1:20, or about 1:25; - a ratio of modifications, substitutions, and / or substitutions of light chain N200 to heavy chain Q58, M64, N68, N70, N77, M80, N93, M103, Q125, N132, Q139, Q275, N4191, Q5216, or N5262 of about 25:1, about 20:1, about 15:1, about 10:1, about 9:1, about 8:1, about 7:1, about 6:1, about 5:1, about 4:1, about 3:1, about 2:1, about 1:1, about 1:2, about 1:3, about 1:4, about 1:5, about 1:6, about 1:7, about 1:8, about 1:9, about 1:10, about 1:15, about 1:20, or about 1:25; - a ratio of modifications, substitutions, and / or substitutions of light chain Q202 to heavy chain Q58, M64, N68, N70, N77, M80, N93, M103, Q125, N132, Q139, Q275, N4191, Q5216, or N5262 of about 25:1, about 20:1, about 15:1, about 10:1, about 9:1, about 8:1, about 7:1, about 6:1, about 5:1, about 4:1, about 3:1, about 2:1, about 1:1, about 1:2, about 1:3, about 1:4, about 1:5, about 1:6, about 1:7, about 1:8, about 1:9, about 1:10, about 1:15, about 1:20, or about 1:25; - a ratio of modifications, substitutions, and / or substitutions of light chain N204 to heavy chain Q58, M64, N68, N70, N77, M80, N93, M103, Q125, N132, Q139, Q275, N4191, Q5216, or N5262 of about 25:1, about 20:1, about 15:1, about 10:1, about 9:1, about 8:1, about 7:1, about 6:1, about 5:1, about 4:1, about 3:1, about 2:1, about 1:1, about 1:2, about 1:3, about 1:4, about 1:5, about 1:6, about 1:7, about 1:8, about 1:9, about 1:10, about 1:15, about 1:20, or about 1:25; - a ratio of modifications, substitutions, and / or substitutions of light chain N240 to heavy chain Q58, M64, N68, N70, N77, M80, N93, M103, Q125, N132, Q139, Q275, N4191, Q5216, or N5262 of about 25:1, about 20:1, about 15:1, about 10:1, about 9:1, about 8:1, about 7:1, about 6:1, about 5:1, about 4:1, about 3:1, about 2:1, about 1:1, about 1:2, about 1:3, about 1:4, about 1:5, about 1:6, about 1:7, about 1:8, about 1:9, about 1:10, about 1:15, about 1:20, or about 1:25; - a ratio of light chain N248 to heavy chain Q58, M64, N68, N70, N77, M80, N93, M103, Q125, N132, Q139, Q275, N4191, Q5216, or N5262 modifications, substitutions, and / or substitutions of about 25:1, about 20:1, about 15:1, about 10:1, about 9:1, about 8:1, about 7:1, about 6:1, about 5:1, about 4:1, about 3:1, about 2:1, about 1:1, about 1:2, about 1:3, about 1:4, about 1:5, about 1:6, about 1:7, about 1:8, about 1:9, about 1:10, about 1:15, about 1:20, or about 1:25; - a ratio of light chain Q255 to heavy chain Q58, M64, N68, N70, N77, M80, N93, M103, Q125, N132, Q139, Q275, N4191, Q5216, or N5262 modifications, substitutions, and / or substitutions of about 25:1, about 20:1, about 15:1, about 10:1, about 9:1, about 8:1, about 7:1, about 6:1, about 5:1, about 4:1, about 3:1, about 2:1, about 1:1, about 1:2, about 1:3, about 1:4, about 1:5, about 1:6, about 1:7, about 1:8, about 1:9, about 1:10, about 1:15, about 1:20, or about 1:25;

[0013] In some embodiments, the present disclosure provides compositions comprising a plurality of peptides or protein fragments, e.g., a plurality of fibroin light chain and p25 peptides or fibroin light chain and p25 fragments, and comprising a plurality of amino acid modifications, substitutions, and / or substitutions, wherein specific positions comprise a ratio of one or more modifications, substitutions, and / or substitutions selected from the following: - a ratio of light chain N23 to p25 Q62, N93, M120, N149, N172, N174, or N202 modifications, substitutions, and / or substitutions of about 25:1, about 20:1, about 15:1, about 10:1, about 9:1, about 8:1, about 7:1, about 6:1, about 5:1, about 4:1, about 3:1, about 2:1, about 1:1, about 1:2, about 1:3, about 1:4, about 1:5, about 1:6, about 1:7, about 1:8, about 1:9, about 1:10, about 1:15, about 1:20, or about 1:25; - a ratio of light chain Q24 to p25 Q62, N93, M120, N149, N172, N174, or N202 modifications, substitutions, and / or substitutions of about 25:1, about 20:1, about 15:1, about 10:1, about 9:1, about 8:1, about 7:1, about 6:1, about 5:1, about 4:1, about 3:1, about 2:1, about 1:1, about 1:2, about 1:3, about 1:4, about 1:5, about 1:6, about 1:7, about 1:8, about 1:9, about 1:10, about 1:15, about 1:20, or about 1:25; - a ratio of light chain N28 to p25 Q62, N93, M120, N149, N172, N174, or N202 modifications, substitutions, and / or substitutions of about 25:1, about 20:1, about 15:1, about 10:1, about 9:1, about 8:1, about 7:1, about 6:1, about 5:1, about 4:1, about 3:1, about 2:1, about 1:1, about 1:2, about 1:3, about 1:4, about 1:5, about 1:6, about 1:7, about 1:8, about 1:9, about 1:10, about 1:15, about 1:20, or about 1:25; - a ratio of light chain M69 to p25 Q62, N93, M120, N149, N172, N174, or N202 modifications, substitutions, and / or substitutions of about 25:1, about 20:1, about 15:1, about 10:1, about 9:1, about 8:1, about 7:1, about 6:1, about 5:1, about 4:1, about 3:1, about 2:1, about 1:1, about 1:2, about 1:3, about 1:4, about 1:5, about 1:6, about 1:7, about 1:8, about 1:9, about 1:10, about 1:15, about 1:20, or about 1:25; - a ratio of light chain N105 to p25 Q62, N93, M120, N149, N172, N174, or N202 modifications, substitutions, and / or substitutions of about 25:1, about 20:1, about 15:1, about 10:1, about 9:1, about 8:1, about 7:1, about 6:1, about 5:1, about 4:1, about 3:1, about 2:1, about 1:1, about 1:2, about 1:3, about 1:4, about 1:5, about 1:6, about 1:7, about 1:8, about 1:9, about 1:10, about 1:15, about 1:20, or about 1:25; - a ratio of light chain N108 to p25 Q62, N93, M120, N149, N172, N174, or N202 modifications, substitutions, and / or substitutions of about 25:1, about 20:1, about 15:1, about 10:1, about 9:1, about 8:1, about 7:1, about 6:1, about 5:1, about 4:1, about 3:1, about 2:1, about 1:1, about 1:2, about 1:3, about 1:4, about 1:5, about 1:6, about 1:7, about 1:8, about 1:9, about 1:10, about 1:15, about 1:20, or about 1:25; - a ratio of light chain N118 to p25 Q62, N93, M120, N149, N172, N174, or N202 modifications, substitutions, and / or substitutions of about 25:1, about 20:1, about 15:1, about 10:1, about 9:1, about 8:1, about 7:1, about 6:1, about 5:1, about 4:1, about 3:1, about 2:1, about 1:1, about 1:2, about 1:3, about 1:4, about 1:5, about 1:6, about 1:7, about 1:8, about 1:9, about 1:10, about 1:15, about 1:20, or about 1:25; - a ratio of light chain N136 to p25 Q62, N93, M120, N149, N172, N174, or N202 modifications, substitutions, and / or substitutions of about 25:1, about 20:1, about 15:1, about 10:1, about 9:1, about 8:1, about 7:1, about 6:1, about 5:1, about 4:1, about 3:1, about 2:1, about 1:1, about 1:2, about 1:3, about 1:4, about 1:5, about 1:6, about 1:7, about 1:8, about 1:9, about 1:10, about 1:15, about 1:20, or about 1:25; - a ratio of light chain N138 to p25 Q62, N93, M120, N149, N172, N174, or N202 modifications, substitutions, and / or substitutions of about 25:1, about 20:1, about 15:1, about 10:1, about 9:1, about 8:1, about 7:1, about 6:1, about 5:1, about 4:1, about 3:1, about 2:1, about 1:1, about 1:2, about 1:3, about 1:4, about 1:5, about 1:6, about 1:7, about 1:8, about 1:9, about 1:10, about 1:15, about 1:20, or about 1:25; - a ratio of light chain Q149 to p25 Q62, N93, M120, N149, N172, N174, or N202 modifications, substitutions, and / or substitutions of about 25:1, about 20:1, about 15:1, about 10:1, about 9:1, about 8:1, about 7:1, about 6:1, about 5:1, about 4:1, about 3:1, about 2:1, about 1:1, about 1:2, about 1:3, about 1:4, about 1:5, about 1:6, about 1:7, about 1:8, about 1:9, about 1:10, about 1:15, about 1:20, or about 1:25; - a ratio of light chain N186 to p25 Q62, N93, M120, N149, N172, N174, or N202 modifications, substitutions, and / or substitutions of about 25:1, about 20:1, about 15:1, about 10:1, about 9:1, about 8:1, about 7:1, about 6:1, about 5:1, about 4:1, about 3:1, about 2:1, about 1:1, about 1:2, about 1:3, about 1:4, about 1:5, about 1:6, about 1:7, about 1:8, about 1:9, about 1:10, about 1:15, about 1:20, or about 1:25; - a ratio of light chain N200 to p25 Q62, N93, M120, N149, N172, N174, or N202 modifications, substitutions, and / or substitutions of about 25:1, about 20:1, about 15:1, about 10:1, about 9:1, about 8:1, about 7:1, about 6:1, about 5:1, about 4:1, about 3:1, about 2:1, about 1:1, about 1:2, about 1:3, about 1:4, about 1:5, about 1:6, about 1:7, about 1:8, about 1:9, about 1:10, about 1:15, about 1:20, or about 1:25; - a ratio of light chain Q202 to p25 Q62, N93, M120, N149, N172, N174, or N202 modifications, substitutions, and / or substitutions of about 25:1, about 20:1, about 15:1, about 10:1, about 9:1, about 8:1, about 7:1, about 6:1, about 5:1, about 4:1, about 3:1, about 2:1, about 1:1, about 1:2, about 1:3, about 1:4, about 1:5, about 1:6, about 1:7, about 1:8, about 1:9, about 1:10, about 1:15, about 1:20, or about 1:25; - a ratio of light chain N204 to p25 Q62, N93, M120, N149, N172, N174, or N202 modifications, substitutions, and / or substitutions of about 25:1, about 20:1, about 15:1, about 10:1, about 9:1, about 8:1, about 7:1, about 6:1, about 5:1, about 4:1, about 3:1, about 2:1, about 1:1, about 1:2, about 1:3, about 1:4, about 1:5, about 1:6, about 1:7, about 1:8, about 1:9, about 1:10, about 1:15, about 1:20, or about 1:25; - a ratio of light chain N240 to p25 Q62, N93, M120, N149, N172, N174, or N202 modifications, substitutions, and / or substitutions of about 25:1, about 20:1, about 15:1, about 10:1, about 9:1, about 8:1, about 7:1, about 6:1, about 5:1, about 4:1, about 3:1, about 2:1, about 1:1, about 1:2, about 1:3, about 1:4, about 1:5, about 1:6, about 1:7, about 1:8, about 1:9, about 1:10, about 1:15, about 1:20, or about 1:25; - a ratio of light chain N248 to p25 Q62, N93, M120, N149, N172, N174, or N202 modifications, substitutions, and / or substitutions of about 25:1, about 20:1, about 15:1, about 10:1, about 9:1, about 8:1, about 7:1, about 6:1, about 5:1, about 4:1, about 3:1, about 2:1, about 1:1, about 1:2, about 1:3, about 1:4, about 1:5, about 1:6, about 1:7, about 1:8, about 1:9, about 1:10, about 1:15, about 1:20, or about 1:25; - a ratio of light chain Q255 to p25 Q62, N93, M120, N149, N172, N174, or N202 modifications, substitutions, and / or substitutions of about 25:1, about 20:1, about 15:1, about 10:1, about 9:1, about 8:1, about 7:1, about 6:1, about 5:1, about 4:1, about 3:1, about 2:1, about 1:1, about 1:2, about 1:3, about 1:4, about 1:5, about 1:6, about 1:7, about 1:8, about 1:9, about 1:10, about 1:15, about 1:20, or about 1:25;

[0014] In some embodiments, the present disclosure provides compositions that include a plurality of peptides or protein fragments, e.g., a plurality of p25 fibroin peptides or fibroin fragments, and that include a plurality of amino acid modifications, substitutions, and / or substitutions, where specific positions include a ratio of one or more modifications, substitutions, and / or substitutions selected from the following: a Q62 to N93 modification, substitution, and / or replacement ratio of about 25:1, about 20:1, about 15:1, about 10:1, about 9:1, about 8:1, about 7:1, about 6:1, about 5:1, about 4:1, about 3:1, about 2:1, about 1:1, about 1:2, about 1:3, about 1:4, about 1:5, about 1:6, about 1:7, about 1:8, about 1:9, about 1:10, about 1:15, about 1:20, or about 1:25; a Q62 to M120 modification, substitution, and / or replacement ratio of about 25:1, about 20:1, about 15:1, about 10:1, about 9:1, about 8:1, about 7:1, about 6:1, about 5:1, about 4:1, about 3:1, about 2:1, about 1:1, about 1:2, about 1:3, about 1:4, about 1:5, about 1:6, about 1:7, about 1:8, about 1:9, about 1:10, about 1:15, about 1:20, or about 1:25; a Q62 to N149 modification, substitution, and / or replacement ratio of about 25:1, about 20:1, about 15:1, about 10:1, about 9:1, about 8:1, about 7:1, about 6:1, about 5:1, about 4:1, about 3:1, about 2:1, about 1:1, about 1:2, about 1:3, about 1:4, about 1:5, about 1:6, about 1:7, about 1:8, about 1:9, about 1:10, about 1:15, about 1:20, or about 1:25; a Q62 to N172 modification, substitution, and / or replacement ratio of about 25:1, about 20:1, about 15:1, about 10:1, about 9:1, about 8:1, about 7:1, about 6:1, about 5:1, about 4:1, about 3:1, about 2:1, about 1:1, about 1:2, about 1:3, about 1:4, about 1:5, about 1:6, about 1:7, about 1:8, about 1:9, about 1:10, about 1:15, about 1:20, or about 1:25; a Q62 to N174 modification, substitution, and / or replacement ratio of about 25:1, about 20:1, about 15:1, about 10:1, about 9:1, about 8:1, about 7:1, about 6:1, about 5:1, about 4:1, about 3:1, about 2:1, about 1:1, about 1:2, about 1:3, about 1:4, about 1:5, about 1:6, about 1:7, about 1:8, about 1:9, about 1:10, about 1:15, about 1:20, or about 1:25; a modification, substitution, and / or replacement ratio of Q62 to N202 of about 25:1, about 20:1, about 15:1, about 10:1, about 9:1, about 8:1, about 7:1, about 6:1, about 5:1, about 4:1, about 3:1, about 2:1, about 1:1, about 1:2, about 1:3, about 1:4, about 1:5, about 1:6, about 1:7, about 1:8, about 1:9, about 1:10, about 1:15, about 1:20, or about 1:25; an N93 to M120 modification, substitution, and / or replacement ratio of about 25:1, about 20:1, about 15:1, about 10:1, about 9:1, about 8:1, about 7:1, about 6:1, about 5:1, about 4:1, about 3:1, about 2:1, about 1:1, about 1:2, about 1:3, about 1:4, about 1:5, about 1:6, about 1:7, about 1:8, about 1:9, about 1:10, about 1:15, about 1:20, or about 1:25; an N93 to N149 modification, substitution, and / or replacement ratio of about 25:1, about 20:1, about 15:1, about 10:1, about 9:1, about 8:1, about 7:1, about 6:1, about 5:1, about 4:1, about 3:1, about 2:1, about 1:1, about 1:2, about 1:3, about 1:4, about 1:5, about 1:6, about 1:7, about 1:8, about 1:9, about 1:10, about 1:15, about 1:20, or about 1:25; an N93 to N172 modification, substitution, and / or replacement ratio of about 25:1, about 20:1, about 15:1, about 10:1, about 9:1, about 8:1, about 7:1, about 6:1, about 5:1, about 4:1, about 3:1, about 2:1, about 1:1, about 1:2, about 1:3, about 1:4, about 1:5, about 1:6, about 1:7, about 1:8, about 1:9, about 1:10, about 1:15, about 1:20, or about 1:25; an N93 to N174 modification, substitution, and / or replacement ratio of about 25:1, about 20:1, about 15:1, about 10:1, about 9:1, about 8:1, about 7:1, about 6:1, about 5:1, about 4:1, about 3:1, about 2:1, about 1:1, about 1:2, about 1:3, about 1:4, about 1:5, about 1:6, about 1:7, about 1:8, about 1:9, about 1:10, about 1:15, about 1:20, or about 1:25; an N93 to N202 modification, substitution, and / or replacement ratio of about 25:1, about 20:1, about 15:1, about 10:1, about 9:1, about 8:1, about 7:1, about 6:1, about 5:1, about 4:1, about 3:1, about 2:1, about 1:1, about 1:2, about 1:3, about 1:4, about 1:5, about 1:6, about 1:7, about 1:8, about 1:9, about 1:10, about 1:15, about 1:20, or about 1:25; - an M120 to N149 modification, substitution, and / or replacement ratio of about 25:1, about 20:1, about 15:1, about 10:1, about 9:1, about 8:1, about 7:1, about 6:1, about 5:1, about 4:1, about 3:1, about 2:1, about 1:1, about 1:2, about 1:3, about 1:4, about 1:5, about 1:6, about 1:7, about 1:8, about 1:9, about 1:10, about 1:15, about 1:20, or about 1:25; - an M120 to N172 modification, substitution, and / or replacement ratio of about 25:1, about 20:1, about 15:1, about 10:1, about 9:1, about 8:1, about 7:1, about 6:1, about 5:1, about 4:1, about 3:1, about 2:1, about 1:1, about 1:2, about 1:3, about 1:4, about 1:5, about 1:6, about 1:7, about 1:8, about 1:9, about 1:10, about 1:15, about 1:20, or about 1:25; - a ratio of M120 to N174 modification, substitution, and / or replacement of about 25:1, about 20:1, about 15:1, about 10:1, about 9:1, about 8:1, about 7:1, about 6:1, about 5:1, about 4:1, about 3:1, about 2:1, about 1:1, about 1:2, about 1:3, about 1:4, about 1:5, about 1:6, about 1:7, about 1:8, about 1:9, about 1:10, about 1:15, about 1:20, or about 1:25; - a ratio of M120 to N202 modification, substitution, and / or replacement of about 25:1, about 20:1, about 15:1, about 10:1, about 9:1, about 8:1, about 7:1, about 6:1, about 5:1, about 4:1, about 3:1, about 2:1, about 1:1, about 1:2, about 1:3, about 1:4, about 1:5, about 1:6, about 1:7, about 1:8, about 1:9, about 1:10, about 1:15, about 1:20, or about 1:25; a N149 to N172 modification, substitution, and / or replacement ratio of about 25:1, about 20:1, about 15:1, about 10:1, about 9:1, about 8:1, about 7:1, about 6:1, about 5:1, about 4:1, about 3:1, about 2:1, about 1:1, about 1:2, about 1:3, about 1:4, about 1:5, about 1:6, about 1:7, about 1:8, about 1:9, about 1:10, about 1:15, about 1:20, or about 1:25; a N149 to N174 modification, substitution, and / or replacement ratio of about 25:1, about 20:1, about 15:1, about 10:1, about 9:1, about 8:1, about 7:1, about 6:1, about 5:1, about 4:1, about 3:1, about 2:1, about 1:1, about 1:2, about 1:3, about 1:4, about 1:5, about 1:6, about 1:7, about 1:8, about 1:9, about 1:10, about 1:15, about 1:20, or about 1:25; a N149 to N202 modification, substitution, and / or replacement ratio of about 25:1, about 20:1, about 15:1, about 10:1, about 9:1, about 8:1, about 7:1, about 6:1, about 5:1, about 4:1, about 3:1, about 2:1, about 1:1, about 1:2, about 1:3, about 1:4, about 1:5, about 1:6, about 1:7, about 1:8, about 1:9, about 1:10, about 1:15, about 1:20, or about 1:25; a N172 to N174 modification, substitution, and / or replacement ratio of about 25:1, about 20:1, about 15:1, about 10:1, about 9:1, about 8:1, about 7:1, about 6:1, about 5:1, about 4:1, about 3:1, about 2:1, about 1:1, about 1:2, about 1:3, about 1:4, about 1:5, about 1:6, about 1:7, about 1:8, about 1:9, about 1:10, about 1:15, about 1:20, or about 1:25; a N172 to N202 modification, substitution, and / or replacement ratio of about 25:1, about 20:1, about 15:1, about 10:1, about 9:1, about 8:1, about 7:1, about 6:1, about 5:1, about 4:1, about 3:1, about 2:1, about 1:1, about 1:2, about 1:3, about 1:4, about 1:5, about 1:6, about 1:7, about 1:8, about 1:9, about 1:10, about 1:15, about 1:20, or about 1:25; a N174 to N202 modification, substitution, and / or replacement ratio of about 25:1, about 20:1, about 15:1, about 10:1, about 9:1, about 8:1, about 7:1, about 6:1, about 5:1, about 4:1, about 3:1, about 2:1, about 1:1, about 1:2, about 1:3, about 1:4, about 1:5, about 1:6, about 1:7, about 1:8, about 1:9, about 1:10, about 1:15, about 1:20, or about 1:25;

[0015] In some embodiments, the present disclosure provides compositions that include a plurality of peptides or protein fragments, e.g., a plurality of p25 and heavy chain peptides or p25 and heavy chain fragments, and that include a plurality of amino acid modifications, substitutions, and / or substitutions, where specific positions include a ratio of one or more modifications, substitutions, and / or substitutions selected from the following: - a ratio of p25 Q62 to heavy chain Q58, M64, N68, N70, N77, M80, N93, M103, Q125, N132, Q139, Q275, N4191, Q5216, or N5262 modifications, substitutions, and / or substitutions of about 25:1, about 20:1, about 15:1, about 10:1, about 9:1, about 8:1, about 7:1, about 6:1, about 5:1, about 4:1, about 3:1, about 2:1, about 1:1, about 1:2, about 1:3, about 1:4, about 1:5, about 1:6, about 1:7, about 1:8, about 1:9, about 1:10, about 1:15, about 1:20, or about 1:25; - a ratio of p25 N93 to heavy chain Q58, M64, N68, N70, N77, M80, N93, M103, Q125, N132, Q139, Q275, N4191, Q5216, or N5262 modifications, substitutions, and / or substitutions of about 25:1, about 20:1, about 15:1, about 10:1, about 9:1, about 8:1, about 7:1, about 6:1, about 5:1, about 4:1, about 3:1, about 2:1, about 1:1, about 1:2, about 1:3, about 1:4, about 1:5, about 1:6, about 1:7, about 1:8, about 1:9, about 1:10, about 1:15, about 1:20, or about 1:25; - a ratio of p25 M120 to heavy chain Q58, M64, N68, N70, N77, M80, N93, M103, Q125, N132, Q139, Q275, N4191, Q5216, or N5262 modifications, substitutions, and / or substitutions of about 25:1, about 20:1, about 15:1, about 10:1, about 9:1, about 8:1, about 7:1, about 6:1, about 5:1, about 4:1, about 3:1, about 2:1, about 1:1, about 1:2, about 1:3, about 1:4, about 1:5, about 1:6, about 1:7, about 1:8, about 1:9, about 1:10, about 1:15, about 1:20, or about 1:25; - a ratio of p25 N149 to heavy chain Q58, M64, N68, N70, N77, M80, N93, M103, Q125, N132, Q139, Q275, N4191, Q5216, or N5262 modifications, substitutions, and / or substitutions of about 25:1, about 20:1, about 15:1, about 10:1, about 9:1, about 8:1, about 7:1, about 6:1, about 5:1, about 4:1, about 3:1, about 2:1, about 1:1, about 1:2, about 1:3, about 1:4, about 1:5, about 1:6, about 1:7, about 1:8, about 1:9, about 1:10, about 1:15, about 1:20, or about 1:25; - a ratio of p25 N172 to heavy chain Q58, M64, N68, N70, N77, M80, N93, M103, Q125, N132, Q139, Q275, N4191, Q5216, or N5262 modifications, substitutions, and / or substitutions of about 25:1, about 20:1, about 15:1, about 10:1, about 9:1, about 8:1, about 7:1, about 6:1, about 5:1, about 4:1, about 3:1, about 2:1, about 1:1, about 1:2, about 1:3, about 1:4, about 1:5, about 1:6, about 1:7, about 1:8, about 1:9, about 1:10, about 1:15, about 1:20, or about 1:25; - a ratio of p25 N174 to heavy chain Q58, M64, N68, N70, N77, M80, N93, M103, Q125, N132, Q139, Q275, N4191, Q5216, or N5262 modifications, substitutions, and / or substitutions of about 25:1, about 20:1, about 15:1, about 10:1, about 9:1, about 8:1, about 7:1, about 6:1, about 5:1, about 4:1, about 3:1, about 2:1, about 1:1, about 1:2, about 1:3, about 1:4, about 1:5, about 1:6, about 1:7, about 1:8, about 1:9, about 1:10, about 1:15, about 1:20, or about 1:25; - a ratio of p25 N202 to heavy chain Q58, M64, N68, N70, N77, M80, N93, M103, Q125, N132, Q139, Q275, N4191, Q5216, or N5262 modifications, substitutions, and / or substitutions of about 25:1, about 20:1, about 15:1, about 10:1, about 9:1, about 8:1, about 7:1, about 6:1, about 5:1, about 4:1, about 3:1, about 2:1, about 1:1, about 1:2, about 1:3, about 1:4, about 1:5, about 1:6, about 1:7, about 1:8, about 1:9, about 1:10, about 1:15, about 1:20, or about 1:25;

[0016] In some embodiments, the present disclosure provides compositions that include a plurality of peptides or protein fragments, e.g., a plurality of p25 and light chain peptides or fibroin light chain and light chain fragments, and that also include a plurality of amino acid modifications, substitutions, and / or substitutions, and further include, at specific positions, ratios of one or more modifications, substitutions, and / or substitutions selected from the following: - a ratio of p25 Q62 to light chain N23, Q24, N28, M69, N105, N108, N118, N136, N138, Q149, N186, N200, Q202, N204, N240, N248 or Q255 modifications, substitutions, and / or substitutions of about 25:1, about 20:1, about 15:1, about 10:1, about 9:1, about 8:1, about 7:1, about 6:1, about 5:1, about 4:1, about 3:1, about 2:1, about 1:1, about 1:2, about 1:3, about 1:4, about 1:5, about 1:6, about 1:7, about 1:8, about 1:9, about 1:10, about 1:15, about 1:20, or about 1:25; - a ratio of p25 N93 to light chain N23, Q24, N28, M69, N105, N108, N118, N136, N138, Q149, N186, N200, Q202, N204, N240, N248 or Q255 modifications, substitutions, and / or substitutions of about 25:1, about 20:1, about 15:1, about 10:1, about 9:1, about 8:1, about 7:1, about 6:1, about 5:1, about 4:1, about 3:1, about 2:1, about 1:1, about 1:2, about 1:3, about 1:4, about 1:5, about 1:6, about 1:7, about 1:8, about 1:9, about 1:10, about 1:15, about 1:20, or about 1:25; - a ratio of p25 M120 to light chain N23, Q24, N28, M69, N105, N108, N118, N136, N138, Q149, N186, N200, Q202, N204, N240, N248 or Q255 modifications, substitutions, and / or substitutions of about 25:1, about 20:1, about 15:1, about 10:1, about 9:1, about 8:1, about 7:1, about 6:1, about 5:1, about 4:1, about 3:1, about 2:1, about 1:1, about 1:2, about 1:3, about 1:4, about 1:5, about 1:6, about 1:7, about 1:8, about 1:9, about 1:10, about 1:15, about 1:20, or about 1:25; - a ratio of p25 N149 to light chain N23, Q24, N28, M69, N105, N108, N118, N136, N138, Q149, N186, N200, Q202, N204, N240, N248 or Q255 modifications, substitutions, and / or substitutions of about 25:1, about 20:1, about 15:1, about 10:1, about 9:1, about 8:1, about 7:1, about 6:1, about 5:1, about 4:1, about 3:1, about 2:1, about 1:1, about 1:2, about 1:3, about 1:4, about 1:5, about 1:6, about 1:7, about 1:8, about 1:9, about 1:10, about 1:15, about 1:20, or about 1:25; - a ratio of p25 N172 to light chain N23, Q24, N28, M69, N105, N108, N118, N136, N138, Q149, N186, N200, Q202, N204, N240, N248 or Q255 modifications, substitutions, and / or substitutions of about 25:1, about 20:1, about 15:1, about 10:1, about 9:1, about 8:1, about 7:1, about 6:1, about 5:1, about 4:1, about 3:1, about 2:1, about 1:1, about 1:2, about 1:3, about 1:4, about 1:5, about 1:6, about 1:7, about 1:8, about 1:9, about 1:10, about 1:15, about 1:20, or about 1:25; - a ratio of p25 N174 to light chain N23, Q24, N28, M69, N105, N108, N118, N136, N138, Q149, N186, N200, Q202, N204, N240, N248 or Q255 modifications, substitutions, and / or substitutions of about 25:1, about 20:1, about 15:1, about 10:1, about 9:1, about 8:1, about 7:1, about 6:1, about 5:1, about 4:1, about 3:1, about 2:1, about 1:1, about 1:2, about 1:3, about 1:4, about 1:5, about 1:6, about 1:7, about 1:8, about 1:9, about 1:10, about 1:15, about 1:20, or about 1:25; - a ratio of p25 N202 to light chain N23, Q24, N28, M69, N105, N108, N118, N136, N138, Q149, N186, N200, Q202, N204, N240, N248 or Q255 modifications, substitutions, and / or substitutions of about 25:1, about 20:1, about 15:1, about 10:1, about 9:1, about 8:1, about 7:1, about 6:1, about 5:1, about 4:1, about 3:1, about 2:1, about 1:1, about 1:2, about 1:3, about 1:4, about 1:5, about 1:6, about 1:7, about 1:8, about 1:9, about 1:10, about 1:15, about 1:20, or about 1:25;

[0017] The present disclosure provides an article comprising a textile and a coating, wherein the coating comprises a surfactant and / or emulsifier system and a surfactant or emulsifier system selected from the group consisting of about 1 kDa to about 5 kDa, about 5 kDa to about 10 kDa, about 6 kDa to about 17 kDa, about 10 kDa to about 15 kDa, about 14 kDa to about 30 kDa, about 15 kDa to about 20 kDa, about 17 kDa to about 39 kDa, about 20 kDa to about 25 kDa, about 25 kDa to about 30 kDa, and about 30 kDa to about 35 kDa. and silk fibroin fragments having an average weight average molecular weight selected from about 35 kDa to about 40 kDa, about 39 kDa to about 54 kDa, about 39 kDa to about 80 kDa, about 40 kDa to about 45 kDa, about 45 kDa to about 50 kDa, about 50 kDa to about 55 kDa, about 55 kDa to about 60 kDa, about 60 kDa to about 100 kDa, or about 80 kDa to about 144 kDa, and a polydispersity ranging from 1 to about 5. In some embodiments, the coating comprises fibroin peptides and / or fibroin fragments disclosed herein, for example, but not limited to, fibroin peptides and / or protein fragments in which at least one of the amino acids has been modified, substituted, or replaced as disclosed herein. In some embodiments, the silk fibroin fragments have a polydispersity of 1 to about 1.5, about 1.5 to about 2.0, about 2.0 to about 2.5, about 2.5 to about 3.0, about 3.0 to about 3.5, about 3.5 to about 4.0, about 4.0 to about 4.5, or about 4.5 to about 5.0. In some embodiments, the silk fibroin fragments have a polydispersity of about 1.5 to about 3.0. In some embodiments, the silk fibroin fragments comprise one or more of low molecular weight silk fibroin fragments and medium molecular weight silk fibroin fragments. In some embodiments, the article further comprises about 0.01% (w / w) to about 10% (w / w) sericin relative to the silk fibroin fragments. In some embodiments, the woven fabric comprises one or more of polyester, polyamide, polyaramid, polytetrafluoroethylene, polyethylene, polypropylene, polyurethane, silicone, a blend of polyurethane and polyethylene glycol, ultra-high molecular weight polyethylene, high performance polyethylene, nylon, LYCRA (a polyester-polyurethane copolymer, also known as SPANDEX and elastomers), or a blend thereof.In some embodiments, the coating further comprises one or more of a wetting agent, an antifoaming agent, an emollient, a wicking agent, and an antimicrobial agent. In some embodiments, the w / w ratio of silk fibroin fragments to surfactant and / or emulsifier system in the coating is about 99:1, about 98:2, about 97:3, about 96:4, about 95:5, about 94:6, about 93:7, about 92:8, about 91:9, about 90:10, about 89:11, about 88:12, about 87:13, about 86:14, about 85:15, about 84:16, about 83:17, about 82:18, about 81:19, about 80:20, or about 91:21. , about 79:21, about 78:22, about 77:23, about 76:24, about 75:25, about 74:26, about 73:27, about 72:28, about 71:29, about 70:30, about 69:31, about 68:32, about 67:33, about 66:34, about 65:35, about 64:36, about 63:37, about 62:38, about 61:39, about 60:40, about 59:41, about 58:42, about 57:43, about 56:44, about 55:45, about 54:46, about 53:47, approx. 52:48, approx. 51:49, approx. 50:50, approx. 49:51, approx. 48:52, approx. 47:53, approx. 46:54, approx. 45:55, approx. 44:56, approx. 43:57, approx. 42:58, approx. 41:59, approx. 40:60, approx. 39:61, approx. 38:62, approx. 37:63, approx. 36:64, approx. 35:65, approx. 34:66, approx. 33:67, approx. 32:68, approx. 31:69, approx. 30:70, approx. 29:71, approx. 28:72, approx. 2 In some embodiments, the w / w ratio of silk fibroin fragments to surfactant and / or emulsifier in the coating is about 1:1, about 1:2, about 1:4, about 1:8, about 1:16, or about 1:32.In some embodiments, the w / w ratio of silk fibroin fragments to surfactant and / or emulsifier system in the coating is about 1:1, about 1:2, about 1:3, about 1:4, about 1:5, about 1:6, about 1:7, about 1:8, about 1:9, about 1:10, about 1:11, about 1:12, about 1:13, about 1:14, about 1:15, about 1:16, about 1:17, about 1:18, about 1:19, about 1:20, about 1:21, about 1:22, about 1:23, about 1:24, about 1:25, about 1:26, about 1:27, about 1:28, about 1:29, about 1:30, about 1:31, or about 1:32. In some embodiments, the surfactant and / or emulsifier system comprises one or more of polyoxyethylene sorbitan monooleate, polyoxyethylene sorbitan trioleate, polyoxyethylene castor oil, and any combination thereof. In some embodiments, the surfactant and / or emulsifier system comprises one or more of polyoxyethylene (10-30) sorbitan monooleate, polyoxyethylene (10-30) sorbitan trioleate, polyoxyethylene (10-50) castor oil, and any combination thereof. In some embodiments, the surfactant and / or emulsifier system comprises one or more of polyoxyethylene (20) sorbitan monooleate, polyoxyethylene (20) sorbitan trioleate, polyoxyethylene (29) castor oil, and any combination thereof. In some embodiments, the surfactant and / or emulsifier system comprises one or more of polyoxyethylene (20) sorbitan monolaurate, polyoxyethylene (20) sorbitan monopalmitate, polyoxyethylene (20) sorbitan monostearate, polyoxyethylene (20) sorbitan tristearate, and any combination thereof. In some embodiments, the surfactant and / or emulsifier system comprises one or more of sorbitan monofatty acid, sorbitan trifatty acid, castor oil, and any combination thereof. In some embodiments, the surfactant and / or emulsifier system comprises one or more of coco glucoside, decyl glucoside, lauryl glucoside, sucrose cocoate, capryl / caprylyl glucoside, caprylyl / capryl glucoside, and any combination thereof.In some embodiments, the surfactant and / or emulsifier system has an HLB of about 11 to about 13.50. In some embodiments, the surfactant and / or emulsifier system has an HLB of about 11 to about 11.50, about 11.50 to about 12, about 12 to about 12.50, about 12.50 to about 13, or about 13 to about 13.50. In some embodiments, the article has improved moisture management compared to a similar article comprising a similar textile but without a coating. In some embodiments, moisture management is evaluated by a water absorption test, a vertical wicking test, or a drying rate test. In some embodiments, the article has improved drape compared to a similar article comprising a similar textile but without a coating. In some embodiments, the article has improved smoothness compared to a similar article comprising a similar textile but without a coating. In some embodiments, the article has improved hand compared to a similar article comprising a similar textile but without a coating. In some embodiments, the article has a lower charge density at a given pH value compared to a similar article comprising a similar textile but without the coating.

[0018] The present disclosure provides methods for producing silk fibroin-coated textiles, the methods comprising applying a solution comprising a surfactant and / or emulsifier system to a textile, applying a silk fibroin fragment solution to the textile, and drying the textile. In some embodiments, the methods include, but are not limited to, the use of modified silk fragments disclosed herein, such as, but not limited to, fibroin peptides and / or protein fragments in which at least one of the amino acids has been modified, substituted, or replaced as disclosed herein. The present disclosure also provides methods for producing silk fibroin-coated textiles, the methods comprising applying a solution comprising a surfactant and / or emulsifier system and silk fibroin fragments to a textile, and drying the textile. In some embodiments, the concentration of the silk fibroin fragments in the solution ranges from 0.01 g / L to about 100 g / L. In some embodiments, the concentration of the surfactant and / or emulsifier system in the solution ranges from 0.01 g / L to about 100 g / L. In some embodiments, the silk fibroin fragments are from about 1 kDa to about 5 kDa, from about 5 kDa to about 10 kDa, from about 6 kDa to about 17 kDa, from about 10 kDa to about 15 kDa, from about 14 kDa to about 30 kDa, from about 15 kDa to about 20 kDa, from about 17 kDa to about 39 kDa, from about 20 kDa to about 25 kDa, from about 25 kDa to about 30 kDa, from about 30 kDa to about 35 kDa, or from about 35 kDa to about 40 kDa. The silk fibroin fragments have an average weight-average molecular weight selected from about Da to about 40 kDa, about 39 kDa to about 54 kDa, about 39 kDa to about 80 kDa, about 40 kDa to about 45 kDa, about 45 kDa to about 50 kDa, about 50 kDa to about 55 kDa, about 55 kDa to about 60 kDa, about 60 kDa to about 100 kDa, or about 80 kDa to about 144 kDa, and a polydispersity ranging from 1 to about 5. In some embodiments, the silk fibroin fragments have a polydispersity ranging from 1 to about 1.5, about 1.5 to about 2.0, about 2.0 to about 2.5, about 2.5 to about 3.0, about 3.0 to about 3.5, about 3.5 to about 4.0, about 4.0 to about 4.5, or about 4.5 to about 5.0. In some embodiments, the silk fibroin fragments have a polydispersity of about 1.5 to about 3.0.In some embodiments, the silk fibroin fragments include one or more of low molecular weight silk fibroin fragments and medium molecular weight silk fibroin fragments. In some embodiments, the solution further includes about 0.01% (w / w) to about 10% (w / w) sericin relative to the silk fibroin fragments. In some embodiments, the textile includes one or more of polyester, polyamide, polyaramid, polytetrafluoroethylene, polyethylene, polypropylene, polyurethane, silicone, a mixture of polyurethane and polyethylene glycol, ultra-high molecular weight polyethylene, high-performance polyethylene, nylon, LYCRA (polyester-polyurethane copolymer, also known as SPANDEX and elastomer), or mixtures thereof. In some embodiments, the solution further includes one or more of a wetting agent, an antifoaming agent, a softener, a wicking agent, and an antimicrobial agent.In some embodiments, the w / w ratio of silk fibroin fragments to surfactant and / or emulsifier system is about 99:1, about 98:2, about 97:3, about 96:4, about 95:5, about 94:6, about 93:7, about 92:8, about 91:9, about 90:10, about 89:11, about 88:12, about 87:13, about 86:14, about 85:15, about 84:16, about 83:17, about 82:18, about 81:19, about 80:20, about 79:2 1, approx. 78:22, approx. 77:23, approx. 76:24, approx. 75:25, approx. 74:26, approx. 73:27, approx. 72:28, approx. 71:29, approx. 70:30, approx. 69:31, approx. 68:32, approx. 67:33, approx. 66:34, approx. 65:35, approx. 64:36, approx. 63:37, approx. 62:38, approx. 61:39, approx. 60:40, approx. 59:41, approx. 58:42, approx. 57:43, approx. 56:44, approx. 55:45, approx. 54:46, approx. 53:4 7, approx. 52:48, approx. 51:49, approx. 50:50, approx. 49:51, approx. 48:52, approx. 47:53, approx. 46:54, approx. 45:55, approx. 44:56, approx. 43:57, approx. 42:58, approx. 41:59, approx. 40:60, approx. 39:61, approx. 38:62, approx. 37:63, approx. 36:64, approx. 35:65, approx. 34:66, approx. 33:67, approx. 32:68, approx. 31:69, approx. 30:70, approx. 29:71, approx. 28:72, approx. 27: In some embodiments, the w / w ratio of silk fibroin fragments to surfactant and / or emulsifier system is about 1:1, about 1:2, about 1:4, about 1:8, about 1:16, about 1:32, about 1:17, about 1:18, about 1:19, about 1:20, about 1:21, about 1:22, about 1:23, about 1:24, about 1:25, about 1:26, about 1:27, about 1:28, about 1:29, about 1:30, about 1:31, about 1:32, about 1:33, about 1:34, about 1:35, about 1:36, about 1:37, about 1:38, about 1:39, about 1:40, about 1:40, about 1:41, about 1:42, about 1:43, about 1:44, about 1:45, about 1:50, about 1:51, about 1:52, about 1:53, about 1:54, about 1:55, about 1:56, about 1:57, about 1:58, about 1:59, about 1:60, about 1:61, about 1:62, about 1:63, about 1:64, about 1:65, about 1:66, about 1:67, about 1:68, about 1:73, about 26:74, about 25:75, about 24:76, about 23:77, about 22:78, about 21:79, about 20:80, about 19:81, about 18:82, about 17:83, about 16:84, about 15:85,In some embodiments, the w / w ratio of silk fibroin fragments to surfactant and / or emulsifier system is about 1:1, about 1:2, about 1:3, about 1:4, about 1:5, about 1:6, about 1:7, about 1:8, about 1:9, about 1:10, about 1:11, about 1:12, about 1:13, about 1:14, about 1:15, about 1:16, about 1:17, about 1:18, about 1:19, about 1:20, about 1:21, about 1:22, about 1:23, about 1:24, about 1:25, about 1:26, about 1:27, about 1:28, about 1:29, about 1:30, about 1:31, or about 1:32. In some embodiments, the surfactant and / or emulsifier system comprises one or more of polyoxyethylene sorbitan monooleate, polyoxyethylene sorbitan trioleate, polyoxyethylene castor oil, and any combination thereof. In some embodiments, the surfactant and / or emulsifier system comprises one or more of polyoxyethylene (10-30) sorbitan monooleate, polyoxyethylene (10-30) sorbitan trioleate, polyoxyethylene (10-50) castor oil, and any combination thereof. In some embodiments, the surfactant and / or emulsifier system comprises one or more of polyoxyethylene (20) sorbitan monooleate, polyoxyethylene (20) sorbitan trioleate, polyoxyethylene (29) castor oil, and any combination thereof. In some embodiments, the surfactant and / or emulsifier system comprises one or more of polyoxyethylene (20) sorbitan monolaurate, polyoxyethylene (20) sorbitan monopalmitate, polyoxyethylene (20) sorbitan monostearate, polyoxyethylene (20) sorbitan tristearate, and any combination thereof. In some embodiments, the surfactant and / or emulsifier system comprises one or more of sorbitan monofatty acid, sorbitan trifatty acid, castor oil, and any combination thereof. In some embodiments, the surfactant and / or emulsifier system comprises one or more of coco glucoside, decyl glucoside, lauryl glucoside, sucrose cocoate, capryl / caprylyl glucoside, caprylyl / capryl glucoside, and any combination thereof.In some embodiments, the surfactant and / or emulsifier system has an HLB of about 11 to about 13.50. In some embodiments, the surfactant and / or emulsifier system has an HLB of about 11 to about 11.50, about 11.50 to about 12, about 12 to about 12.50, about 12.50 to about 13, or about 13 to about 13.50. In some embodiments, drying includes heating. In some embodiments, the pH of the solution is acidic. In some embodiments, the pH of the solution is about 3.5 to about 4, about 4 to about 4.5, about 4.5 to about 5, about 5 to about 5.5, or about 5.5 to about 6.

[0019] The present disclosure also provides articles prepared by the methods described herein, including, but not limited to, by using modified silk fragments disclosed herein, such as, but not limited to, fibroin peptides and / or protein fragments in which at least one of the amino acids has been modified, substituted, or replaced as disclosed herein. In some embodiments, the articles have improved moisture management compared to a similar article comprising a similar textile but without a coating. In some embodiments, moisture management is evaluated by a water absorption test, a vertical wicking test, or a drying rate test. In some embodiments, the articles have improved drape compared to a similar article comprising a similar textile but without a coating. In some embodiments, the articles have improved smoothness compared to a similar article comprising a similar textile but without a coating. In some embodiments, the articles have improved hand compared to a similar article comprising a similar textile but without a coating. In some embodiments, the articles have a lower charge density at a given pH value compared to a similar article comprising a similar textile but without a coating.

[0020] In some embodiments, the present disclosure provides a method for producing a medicament comprising administering to a subject a medicament of the present invention a medicament of the present invention, comprising administering to a subject a medicament of the present invention a medicament of the present invention, the medicament of the present invention being an medicament of the present invention, the medicament of the present invention being an medicament of the present invention, Further provided are personal care and / or cosmetic formulations and / or compositions, including silk fibroin fragments having, but not limited to, an average weight average molecular weight selected from about 80 kDa to about 144 kDa and a polydispersity of 1 to about 5, and which may also include modified silk protein fragments disclosed herein, e.g., fibroin peptides and / or protein fragments in which at least one of the amino acids has been modified, substituted, or replaced as disclosed herein. In some embodiments, the composition further comprises 0 to 500 ppm lithium bromide. In some embodiments, the composition further comprises 0 to 500 ppm sodium carbonate. In some embodiments, the silk fibroin fragments have a polydispersity of 1 to about 1.5. In some embodiments, the silk fibroin fragments have a polydispersity of about 1.5 to about 2.0. In some embodiments, the silk fibroin fragments have a polydispersity of about 1.5 to about 3.0. In some embodiments, the silk fibroin fragments have a polydispersity of about 2.0 to about 2.5. In some embodiments, the silk fibroin fragments have a polydispersity of about 2.5 to about 3.0. In some embodiments, the silk fibroin fragments are present in the composition at about 0.001% to about 10.0% by weight, based on the total weight of the composition. In some embodiments, the composition further comprises about 0.001% (w / w) to about 10% (w / w) sericin, based on the total weight of the composition. In some embodiments, the composition further comprises about 0.001% (w / w) to about 10% (w / w) sericin, based on the total weight of the silk fibroin fragments.In some embodiments, the silk fibroin fragments do not spontaneously or gradually gel, and do not visually change in color or turbidity, when in aqueous solution for at least 10 days prior to incorporation into the composition. In some embodiments, the silk fibroin fragments are present in the composition at about 0.01 wt % to about 10.0 wt %, based on the total weight of the composition. In some embodiments, the silk fibroin fragments are present in the composition at about 0.01 wt % to about 1.0 wt %, based on the total weight of the composition. In some embodiments, the silk fibroin fragments are present in the composition at about 1.0 wt % to about 2.0 wt %, based on the total weight of the composition. In some embodiments, the silk fibroin fragments are present in the composition at about 2.0 wt % to about 3.0 wt %, based on the total weight of the composition. In some embodiments, the silk fibroin fragments are present in the composition at about 3.0 wt % to about 4.0 wt %, based on the total weight of the composition. In some embodiments, the silk fibroin fragments are present in the composition at about 4.0 wt % to about 5.0 wt %, based on the total weight of the composition. In some embodiments, the silk fibroin fragments are present in the composition at about 5.0% to about 6.0% by weight, based on the total weight of the composition.

[0021] In some embodiments, the composition is formulated as a topical composition. In some embodiments, the composition further comprises a pharmaceutically acceptable carrier. In some embodiments, the composition further comprises a dermatologically acceptable carrier. In some embodiments, the pharmaceutically acceptable carrier comprises one or more of a suspension, emulsion, powder, solution, dispersion, or elixir. In some embodiments, the pharmaceutically acceptable carrier comprises or is formulated as one or more of a gel, jelly, cream, lotion, foam, slurry, ointment, oil, paste, suppository, spray, semi-solid composition, solid composition, stick, or mousse. In some embodiments, the pharmaceutically acceptable carrier comprises one or more of sesame oil, corn oil, cottonseed oil, or peanut oil. In some embodiments, the pharmaceutically acceptable carrier comprises one or more of mannitol or dextrose. In some embodiments, the pharmaceutically acceptable carrier comprises free hyaluronic acid and / or non-crosslinked hyaluronic acid. In some embodiments, the pharmaceutically acceptable carrier comprises one or more of an aliphatic oil, a fatty alcohol, a fatty acid, a glyceride, an acylglycerol, and a phospholipid. In some embodiments, the pharmaceutically acceptable carrier comprises one or more of a monoglyceride, a diglyceride, or a triglyceride. In some embodiments, the pharmaceutically acceptable carrier comprises an aqueous phase. In some embodiments, the pharmaceutically acceptable carrier comprises an oil-in-water emulsion or a water-in-oil emulsion. In some embodiments, the pharmaceutically acceptable carrier comprises one or more of a hydrocarbon oil, a fatty acid, a fatty oil, a fatty acid ester, or a cationic quaternary ammonium salt.

[0022] The presently disclosed embodiments will be further described with reference to the accompanying drawings, in which the drawings are not necessarily to scale, emphasis instead generally being placed upon illustrating the principles of the presently disclosed embodiments. [Brief explanation of the drawings]

[0023] [Figure 1]Ion Exchange Fractionation Scheme for Isolation of Populations Constituting Low-Skid and Medium-Skid Silk / Modified Polypeptide Compositions Low- and medium-skid silk / modified polypeptide compositions contain silk / modified polypeptides that are negatively or positively charged, or neutral. Q Anion Exchange Chromatography (A) was used to isolate these populations. [Figure 2] Chromatogram of low-skid silk / modified polypeptide composition loaded onto a Q-Sepharose HP column (Cytiva). The flow-through contains silk / modified polypeptides that were not trapped within the column and lack negatively charged amino acids. After loading the column with low-skid or medium-skid silk / modified polypeptide composition and collecting the flow-through, the column is washed until the UV-280 absorbance is below 200 AU. The trapped negatively charged silk / modified polypeptides are eluted with a high salt concentration (1 M NaCl) and comprise AS11 and AS22. Chromatography is performed in a Tris-containing buffer, but the flow-through and Q eluate are finally dialyzed in water. [Figure 3] Analytical size exclusion chromatography of low-skid silk, medium-skid silk / modified silk compositions and their constituent AS compositions. Average molecular weight and polydispersity measurements in kDa are shown. [Figure 4] Figure 4A-4B Analytical size exclusion chromatography of low-skid and medium-skid silk / modified peptide compositions and their constituents (see Table 1 for details). Figure 4A: Molecular weights of the various activated silk novel compositions described in this study. Figure 4B: Polydispersity (PDI) of the various activated silk novel compositions described in this study. AS24 reconstitutes the average molecular weight and polydispersity of the low-skid silk / modified peptide composition and is composed of 50% AS12 and 50% AS22 (see Table 1 for details). AS6 reconstitutes the average molecular weight and polydispersity of the medium-skid silk / modified peptide composition and is composed of 50% AS1 and 50% AS11 (see Table 1 for details). [Figure 5]Isoelectric focusing of low-skid silk / modified polypeptide compositions. Lanes 2 and 7: Low-skid silk loaded in different amounts. Lanes 3, 5, 8, and 10: AS12 silk loaded in different amounts in different preparations. Lanes 4, 6, 9, and 11: AS22 silk loaded in different amounts in different preparations. [Figure 6] Self-assembly reactions of low-skid and medium-skid silk / modified peptide compositions and their constituents (see Table 1 for details). Both graphs illustrate the kinetic parameters of gel formation during silk self-assembly. Graph A shows the calculation of three self-assembly kinetic parameters: t, A, and SARF. See text for details. [Figure 7] Self-assembly kinetics of low-skid and medium-skid silk / modified peptide compositions and their constituents (see Table 1 for details). Figure 7A: "Self-assembly rate coefficient" indicates how fast the self-assembly reaction proceeds once the self-assembly reaction is initiated and the self-assembly nuclei are assembled. Figure 7B: "Maximum gel yield" indicates the density of the silk gel after self-assembly is complete. Figure 7C: "Time" is the time required for the self-assembly reaction to produce half of the maximum gel volume. [Figure 8] Low-skid and medium-skid silk / modified peptide compositions and their constituents (see Table 1 for details). The "self-assembly coefficient" reflects the average tendency of silk to self-assemble to form a gel. The above-identified figures illustrate embodiments disclosed herein; however, other embodiments are contemplated, as noted in the discussion. This disclosure presents exemplary embodiments that are representative and not limiting. Numerous other modifications and embodiments may be derived by those skilled in the art that fall within the scope and spirit of the principles of the presently disclosed embodiments. [Figure 9] Figure 9 is a graph of the weight average molecular weight (i.e., average molecular weight or average MW) using size exclusion chromatography with refractive index detector (SEC-RI) plotted as a function of time for solubilized fibroin in 9.3 M LiBr at 100°C to 103°C (i.e., a temperature gradient of 3 degrees Celsius between 100°C and 103°C). [Figure 10]Figure 10 is a graph of the weight average molecular weight (i.e., average molecular weight average or average MW) using size exclusion chromatography with refractive index detector (SEC-RI) plotted as a function of time for solubilized fibroin in 9.3 M LiBr at 122°C to 125°C (i.e., a temperature gradient of 3 degrees Celsius between 122°C and 125°C). [Figure 11] FIG. 11 is a graph illustrating the percentage of amino acid modifications in silk. [Figure 12] Figures 12A-12C are graphs illustrating the proportion of amino acid modifications in low- and medium-skid silks. Figure 12A illustrates heavy chain modifications, Figure 12B illustrates light chain modifications, and Figure 12C illustrates fibrohexamerin modifications. N is asparagine, which becomes aspartic acid, and Q is glutamine, which is deamidated. M corresponds to methionine, which is oxidized. The number after each amino acid indicates its position along the amino acid chain from the corresponding protein. [Figure 13] Figures 13A-13B are graphs illustrating the percentage of amino acid modifications produced and lyophilized in low- and medium-skid silk. Figure 13A illustrates heavy chain modifications, and Figure 13B illustrates light chain modifications. N is asparagine, which becomes aspartic acid, and Q is glutamine, which is deamidated. M corresponds to methionine, which is oxidized. The number after each amino acid indicates its position along the amino acid chain from the corresponding protein. [Figure 14] Figures 14A-14B are graphs illustrating the percentage of amino acid modifications in low-skid silk produced at Walpole and Medford using the skid process with different process parameters and varying levels. Figure 14A illustrates heavy chain modifications, and Figure 14B illustrates light chain modifications. N is asparagine, which becomes aspartic acid, and Q is glutamine, which is deamidated. M corresponds to methionine, which is oxidized. The number after each amino acid indicates its position along the amino acid chain from the corresponding protein. [Figure 15-1]Figures 15A-15D are graphs illustrating the percentage of amino acid modifications produced in low and medium silk by the skid and benchtop processes. N is asparagine, which becomes aspartic acid; Q is glutamine, which is deamidated; and M corresponds to methionine, which is oxidized. The numbers after each amino acid indicate its position along the amino acid chain from the corresponding protein. [Figure 15-2] Continued from Figure 15-1. [Figure 16] FIG. 16 is an illustration of the method used to calculate the percentage ratio of modified amino acids at specific locations along the sequence of each peptide. [Figure 17] Figure 17 illustrates the anion exchange and size exclusion chromatography scheme for the isolation of low-skid silk / modified peptide compositions. Low-skid silk / modified polypeptide compositions are composed of a diverse population of peptides with a wide range of sizes and charges. Using Q-Sepharose anion exchange chromatography as the first step and HiLoad Superdex 200 size exclusion chromatography as the second purification step, distinct populations of low-skid silk / modified polypeptide compositions were separated. The Q-Sepharose eluate was loaded onto HiLoad Superdex 200 size exclusion chromatography, which resulted in size-fractionated negatively charged silk compositions / modified peptides. [Figure 18]Figures 18A and 18B show chromatograms of anion exchange chromatography of the low-skid silk / modified polypeptide composition and subsequent size-exclusion chromatography of the eluate (Q eluate). Figure 18A: Anion exchange chromatography was performed using a Q-Sepharose column (Cytiva). The low-skid silk / modified peptide composition was separated by anion exchange chromatography into an uncharged peptide population (flow-through—light blue background) and the eluted negatively charged silk composition (eluate—light pink background). The light yellow background represents a column wash with 50 mM Tris pH=8.0 prior to elution of the charged peptide population. Figure 18B: The negatively charged eluate was loaded onto a Superdex 200 column and run through the column using 50 mM Tris, 200 mM CaCl2, pH=8.0. When UV-280 absorbance began to increase for the fractions, the low-skid silk / modified peptide composition was collected and separated by size. The relative elution volumes of silk compositions AS77 and AS81 are depicted on the chromatogram. [Figure 19] Figures 19A and 19B illustrate analytical size exclusion chromatography of low-skid silk / modified silk compositions and their constituent AS compositions. Figure 19A: Average molecular weights in kDa of low-skid silk (LS) and AS77-AS81. Figure 19B: Polydispersity (PDI) measurements. Numerical data are presented in Table 7. [Figure 20] Figure 20 is an SDS polyacrylamide gel electrophoresis of low-skid silk / modified polypeptide compositions. Lanes are represented by fraction number in order of elution from a Superdex 200 column and their respective silk compositions: fraction 6 is AS77, fraction 7 is AS78, fraction 8 is AS79, fraction 9 is AS80, and fraction 10 is AS81. [Figure 21]Figures 21A and 21B are graphs illustrating the self-assembly reaction of a low-skid silk / modified peptide composition. A medium-skid silk reaction was used as a positive control. Figure 21A illustrates the kinetic parameters of gel formation during silk self-assembly. Self-assembly parameters for medium-skid silk: Amax is 0.6780 (Abs), SARF is 8.676, T0.5 is 3.668 hours, and FSAF is 3.08 (Abs / min). Figure 21B is a snapshot of the same self-assembly assay at a later time point, 12 days after the assay was set up. None of the fractions tested self-assembled over time. [Figure 22] Figures 22A and 22B illustrate the characterization of low-skid silk compositions by dynamic light scattering. The low-skid silk / modified peptide compositions were diluted to a concentration of 1 mg / mL, filtered, and analyzed by Zetasizer Pro to estimate the diameter particle size of each silk composition. Figure 22A illustrates the intensity diameter particle size distribution measured for silk compositions AS77, AS78, AS79, AS80, and AS81. Figure 22B illustrates the correlogram functions of silk compositions AS77, AS78, AS79, AS80, and AS81. [Figure 23] Figure 23 illustrates a size-exclusion chromatography scheme for the isolation of low-skid silk / modified peptide compositions. The low-skid silk / modified polypeptide compositions were composed of various peptide populations across a wide range of sizes. Using HiLoad Superdex 200 size-exclusion chromatography, distinct populations of low-skid silk / modified polypeptide compositions were separated. [Figure 24] Figure 24 shows a chromatogram of a low-skid silk / modified polypeptide composition loaded onto a Superdex 200 gel filtration column. The low-skid silk / modified peptide composition was loaded onto a Superdex 200 column and run through the column using 50 mM Tris, 200 mM CaCl2, pH = 8.0. When the UV-280 absorbance of the fractions began to increase, the low-skid silk / modified peptide composition was collected and separated by size. The relative elution volumes of silk compositions AS82, AS86, and AS87 are shown on the chromatogram. [Figure 25]Figures 25A and 25B illustrate analytical size exclusion chromatography of low-skid silk / modified silk compositions and their constituent AS compositions. Figure 25A illustrates the average molecular weights in kDa of low-skid silk (LS) and AS82-AS89. Figure 25B shows polydispersity (PDI) measurements. Numerical data are presented in Table 9. [Figure 26] Figure 26 is an SDS polyacrylamide gel electrophoresis of low-skid silk / modified polypeptide compositions. Lanes are represented by fraction number in order of elution from a Superdex 200 column and their respective silk compositions: fraction 6 is AS82, fraction 7 is AS83, fraction 8 is AS84, fraction 9 is AS85, and fraction 10 is AS86. [Figure 27] Figures 27A and 27B are graphs illustrating the self-assembly reaction of low-skid silk / modified peptide compositions. A medium-skid silk reaction was used as a positive control. Figure 27A illustrates the kinetic parameters of gel formation during silk self-assembly. Self-assembly parameters for medium-skid silk: Amax is 0.6978 (Abs), SARF is 8.591, T0.5 is 3.361 hours, and FSAF is 3.46 (Abs / min). Figure 27B is a snapshot of the same self-assembly assay at a later time point, 18 days after the assay was set up. AS87, AS88, and AS89 demonstrate gel formation, which was already observed 5 days into the assay (LS, low-skid silk; MS, medium-skid silk). [Figure 28]Figures 28A-28C are graphs showing the characterization of low-skid silk compositions by dynamic light scattering. Low-skid silk / modified peptide compositions were diluted to a concentration of 1 mg / mL, filtered, and analyzed using a Zetasizer Pro to estimate the particle size of each silk composition. Figure 28A shows the intensity particle size distribution measured for silk compositions AS82, AS83, AS84, AS85, AS86, AS87, AS88, and AS89. Figure 28B shows the intensity particle size distribution measured for silk composition AS82, the low-skid silk / modified peptide composition (LS), and the medium-skid silk / modified peptide composition (MS). Figure 28C shows the correlogram functions for silk compositions AS82, AS83, AS84, AS85, AS86, AS87, AS88, AS89, the low-skid silk / modified peptide composition (LS), and the medium-skid silk / modified peptide composition (MS). [Figure 29]Figure 29 shows the anion exchange chromatography (Q), hydrophobic interaction chromatography (HIC), and size exclusion chromatography (SEC) schemes for the isolation of low-skid silk / modified peptide compositions. Low-skid silk / modified polypeptide compositions are composed of a diverse population of peptides with a wide range of sizes and charges. Separate populations of low-skid silk / modified polypeptide compositions were isolated using Q-Sepharose anion exchange chromatography as the first step, Butyl ImpRes hydrophobic interaction resin as the second step, and HiLoad Superdex 200 size exclusion chromatography as the third purification step. The Q-Sepharose eluate was loaded onto a Butyl ImpRes (HIC) column, and the HIC eluate was loaded onto HiLoad Superdex 200 size exclusion chromatography, which resulted in the fractionation of negatively charged silk compositions / modified peptides, sized according to their hydrophobicity. The Q-Sepharose eluate contained negatively charged peptides of all sizes. These peptides were resolved on a Butyl ImpRes column, resulting in the elution of high molecular weight, negatively charged, slightly hydrophobic silk components / modified peptides. Smaller, negatively charged peptides washed through and did not bind to the Butyl ImpRes column. The Q-HIC (elution) was loaded onto a Superdex 200 and separated by size. [Figure 30-1]Figures 30A-30E show chromatograms of anion exchange, hydrophobic interaction, and size exclusion chromatography of a low-skid silk / modified polypeptide composition. Figure 30A illustrates that anion exchange chromatography was performed using a Q-Sepharose column. The low-skid silk / modified peptide composition was separated by anion exchange chromatography into an uncharged peptide population (flow-through—light blue background) and the eluted negatively charged silk composition (eluate—light pink background). The light yellow background represents a column wash with 50 mM Tris pH 8.0 prior to elution of the charged peptide population. Figure 30B illustrates that the negatively charged eluate (Q eluate) was loaded onto a Butyl ImpRes column in the presence of 300 mM ammonium sulfate [(NH4)2SO4], exposing the hydrophobic domains of the silk peptides, which allow binding to the column. The highly charged peptide population did not bind to the column (flow-through) and is highlighted in light blue. The column was washed until the OD280 decreased to approximately 100 units (light yellow). The bound silk peptides (Q-HIC (elution)) were then eluted using 50 mM Tris, pH 8.0, without ammonium sulfate (light pink). Figure 30C illustrates that Q-HIC (elution) was further fractionated by size exclusion chromatography (SEC) using a Superdex 200 gel filtration column. The Q-HIC (elution) fraction was run through the column using 50 mM Tris, 200 mM CaCl2, pH 8.0. When the UV-280 absorbance of the fractions began to increase, the low-skid silk / modified peptide compositions were collected and separated by size. The relative elution volumes of silk compositions AS90 and AS94 are shown on the chromatogram. Figure 30D illustrates that the Q-HIC (flow-through) fraction was further fractionated by SEC using a Superdex 200 column, following the same procedure as in (VC). The relative elution volumes of silk compositions AS95 and AS100 are depicted on the chromatogram. Figure 30E: Illustrates the overlay of chromatograms (VC) and (VD).The Q-HIC (elution) fraction has a higher molecular weight range and a lower molecular weight range compared to the Q-HIC (flow-through) fraction that elutes later in the SEC. [Figure 30-2] Continued from Figure 30-1. [Figure 30-3] Continued from Figure 30-2. [Figure 31-1] Figures 31A-31B are graphs showing analytical size exclusion chromatography of low-skid silk / modified silk compositions and their constituent AS compositions. Figure 31A: Average molecular weights in kDa of low-skid silk (LS) and AS90-AS100. Figure 31B: Polydispersity (PDI) measurements. Numerical data are presented in Table 11. [Figure 31-2] Continued from Figure 31-1. [Figure 32] Figures 32A-32B are SDS polyacrylamide gel electrophoresis of low-skid silk / modified polypeptide compositions. Figure 32A: Q-HIC (elution) SEC fractions. Figure 32B: Q-HIC (flow-through) SEC fractions. Lanes are represented by fraction number and their respective silk compositions in order of elution from the Superdex 200 column: in Figure 32A, fraction 6 is AS90, fraction 7 is AS91, fraction 8 is AS92, fraction 9 is AS93, and fraction 10 is AS94. In Figure 32B, fraction 8 is AS95, fraction 9 is AS96, fraction 10 is AS97, fraction 11 is AS98, fraction 12 is AS99, and fraction 13 is AS100. [Figure 33] Figure 33 illustrates the self-assembly reaction of a low-skid silk / modified peptide composition. A medium-skid silk reaction was used as a positive control. Kinetic parameters of gel formation during silk self-assembly. Q-HIC (elution) is the elution fraction eluted from the Butyl ImpRes column before SEC purification. LS, low-skid silk; MS, medium-skid silk. Self-assembly parameters for medium-skid silk: Amax is 0.6974 (Abs), SARF is 8.661, T0.5 is 3.834 h, and FSAF is 3.03 (Abs / min). [Figure 34-1]Figures 34A-34F illustrate the characterization of low-skid silk compositions by dynamic light scattering. Low-skid and medium-skid silk / modified peptide compositions were diluted to a concentration of 1 mg / mL, filtered, and analyzed using a Zetasizer Pro to estimate the diameter particle size of each silk composition. Figure 34A: Intensity diameter particle size distributions measured for silk composition AS90, the Q-HIC (elution) fraction (before fractionation by SEC), low-skid silk (LS), and medium-skid silk (MS). Figure 34B: Correlogram functions of the silk compositions presented in (34A). Figure 34C: Intensity diameter particle size distributions measured for silk compositions AS90-AS94 derived from the Q-HIC (elution)-SEC fractionation process. Figure 34D: Correlogram functions of the silk compositions presented in (34C). Figure 34E: Intensity diameter particle size distributions measured for silk compositions AS95-AS100 derived from the Q-HIC (flow-through)-SEC fractionation process. Figure 34F: Correlogram function of the silk composition presented in (34E). [Figure 34-2] Continued from Figure 34-1. [Figure 34-3] Continued from Figure 34-2. [Figure 35] 1 illustrates a size-exclusion chromatography scheme for the isolation of a medium-skid silk / modified peptide composition. The medium-skid silk / modified polypeptide composition is composed of various peptide populations across a wide range of sizes. Using HiLoad Superdex 200 size-exclusion chromatography, distinct populations of the medium-skid silk / modified polypeptide composition could be separated. [Figure 36] This is a chromatogram of a medium-skid silk / modified polypeptide composition loaded onto a Superdex 200 gel filtration column. The medium-skid silk / modified peptide composition was loaded onto a Superdex 200 column and run through the column using 50 mM Tris, 200 mM CaCl2, pH = 8.0. The medium-skid silk / modified peptide composition was collected and separated by size when the UV-280 absorbance began to increase for the fractions. The relative elution volumes of silk compositions AS107 and AS111 are shown on the chromatogram. [Figure 37]Analytical size exclusion chromatography of medium-skid silk, medium-skid silk / modified silk compositions, and their constituent AS compositions are illustrated in Figure 37A. Average molecular weights in kDa of medium-skid silk (MS) and AS106-AS111 are shown. Figure 37B. Polydispersity (PDI) measurements are shown. Numerical data are presented in Table 14. [Figure 38] SDS polyacrylamide gel electrophoresis of medium-skid silk / modified polypeptide compositions. Lanes are represented by fraction number in order of elution from a Superdex 200 column and their respective silk compositions: fraction 6 is AS107, fraction 7 is AS108, fraction 8 is AS109, fraction 9 is AS110, and fraction 10 is AS111. [Figure 39] Figure 1 illustrates the self-assembly reaction of medium-skid silk / modified peptide compositions. Kinetic parameters of gel formation during silk self-assembly. The red dashed line shows how the self-assembly parameters Amax, SARF, and T0.5 were calculated for unfractionated medium-skid silk (MS). These numerically calculated parameters for silk compositions AS106-AS111 can be found in Table 16. Low-skid silk (LS) was used as a negative control. LS, low-skid silk; MS, medium-skid silk. [Figure 40] Figure 40 illustrates the characterization of medium-skid silk compositions by dynamic light scattering. The medium-skid silk / modified peptide compositions were diluted to a concentration of 1 mg / mL, filtered, and analyzed by Zetasizer Pro to estimate the particle size of each silk composition. Figure 40A: Intensity particle size distribution measured for silk compositions AS106, AS107, AS108, AS109, AS110, AS111, and medium-skid (MS). Figure 50B: Correlation functions for the silk compositions presented in (40A). [Figure 41]Figure 1 illustrates an anion exchange and size exclusion chromatography scheme for the isolation of medium-skid silk / modified peptide compositions. The medium-skid silk / modified polypeptide composition is composed of a diverse population of peptides with a wide range of sizes and charges. Using Q-Sepharose anion exchange chromatography as the first step and HiLoad Superdex 200 size exclusion chromatography as the second purification step, distinct populations of the medium-skid silk / modified polypeptide composition were separated. The Q-Sepharose eluate was loaded onto HiLoad Superdex 200 size exclusion chromatography, which resulted in size-fractionated negatively charged silk compositions / modified peptides. [Figure 42] Chromatograms of anion exchange chromatography of the medium-skid silk / modified polypeptide composition and subsequent size-exclusion chromatography of the eluate (Q eluate). Figure 42A: Anion exchange chromatography was performed using a Q-Sepharose column (Cytiva). The medium-skid silk / modified peptide composition was separated by anion exchange chromatography into an uncharged peptide population (flow-through—light blue background) and the eluted negatively charged silk composition (eluate—light pink background). The light yellow background represents a column wash with 50 mM Tris pH=8.0 prior to elution of the charged peptide population. Figure 42B: The negatively charged eluate (Q eluate) was loaded onto a Superdex 200 column and run through the column using 50 mM Tris, 200 mM CaCl2, pH=8.0. When UV-280 absorbance began to increase for the fractions, the medium-skid silk / modified peptide composition was collected and separated by size. The relative elution volumes of silk compositions AS101 and AS105 are depicted on the chromatogram. [Figure 43] Analytical size exclusion chromatography of medium-skid silk, medium-skid silk / modified silk compositions, and their constituent AS compositions are illustrated in Figure 43A. Average molecular weights in kDa of medium-skid silk (MS) and AS101-AS105 are shown. Figure 43B. Polydispersity (PDI) measurements are shown. Numerical data are presented in Table 16. [Figure 44A]SDS polyacrylamide gel electrophoresis of medium-skid silk / modified polypeptide compositions. Lanes are represented by fraction number in order of elution from a Superdex 200 column and their respective silk compositions: fraction 6 is AS101, fraction 7 is AS102, fraction 8 is AS103, fraction 9 is AS104, and fraction 10 is AS105. [Figure 44B] Continuation of Figure 44A. Figure 44B illustrates the self-assembly reaction of a medium-skid silk / modified peptide composition. A low-skid silk reaction was used as a negative control. Kinetic parameters of gel formation during silk self-assembly are shown. The red dotted lines are shown for clarity in the calculation of the Amax, SARF (self-assembly rate factor), and T0.5 parameters in Table 17. [Figure 45-1] 45A-45C are graphs illustrating the characterization of medium-skid silk compositions by dynamic light scattering. The medium-skid silk / modified peptide compositions were diluted to a concentration of 1 mg / mL, filtered, and analyzed by a Zetasizer Pro (Malvern) to estimate the diameter particle size of each silk composition. Figure 45A: Intensity diameter particle size distribution by intensity measured for silk compositions AS101, AS102, AS103, AS104, and AS105. Figure 45B: Intensity diameter particle size distribution by intensity measured for silk compositions AS101, AS105, and medium-skid silk (MS) to highlight the size difference between AS101 and AS105. Figure 45C: Correlogram functions for silk compositions AS101, AS102, AS103, AS104, AS105, and medium-skid silk (MS). [Figure 45-2] Continued from Figure 45-1. [Figure 46] An example of the values ​​of the three molar mass moments (Mn, Mw, and Mz), which relate to the molar mass and the number of molecules at each molar mass. This example is applicable to polydisperse samples; for monodisperse samples, Mn = Mw = Mz. [Figure 47]Figures 47A-B are analytical SEC-MALS of low, medium, and high molecular weight silks. Figure 47A: Weight average molecular weight in kDa for low, medium, and high molecular weight silks. Figure 47B: Polydispersity index (PDI) measurements for low, medium, and high molecular weight silks. [Figure 48] Analytical SEC-MALS of low, medium, and high molecular weight silks comprised of silk types produced by different process parameters and variable levels. Individual data points are shown, and the mean is represented by the height of the box. Bars encompass one standard deviation. Figure 48A: Weight average molecular weight ranges for low, medium, and high molecular weight silks. Figure 48B: PDI ranges for low, medium, and high molecular weight silks. [Figure 49] Figures 49A-B are analytical SEC-MALS of low-skid silk / modified silk compositions and constituent AS compositions separated by Q-SEC (Q eluent). Figure 49A: Average molecular weights in kDa of low-skid silk (LS) and AS77-AS81. Figure 49B: Polydispersity (PDI) measurements are shown. Numerical data are presented in Table 24. [Figure 50] Figures 50A-50B are analytical SEC-MALS of low-skid silk / modified silk compositions and constituent AS compositions separated by SEC. Figure 50A: Average molecular weights in kDa of low-skid silk (LS) and AS82-AS89. Figure 50B: Polydispersity (PDI) measurements. Numerical data are presented in Table 25. [Figure 51] Figures 51A-B are analytical SEC-MALS of low-skid silk / modified silk compositions and constituent AS compositions separated by Q-HIC-SEC (Q-HIC-eluent). Figure 51A: Average molecular weights in kDa of low-skid silk (LS) and AS90-AS94. Figure 51B: Polydispersity (PDI) measurements. Numerical data are presented in Table 26. [Figure 52]Figures 52A-B are analytical SEC-MALS of low-skid silk / modified silk compositions and constituent AS compositions separated by Q-HIC-SEC (Q-HIC-flow-through). Figure 52A: Average molecular weights in kDa of low-skid silk (LS) and AS95-AS100. Figure 52B: Polydispersity (PDI) measurements. Numerical data are presented in Table 26. [Figure 53] Figures 53A-B are analytical SEC-MALS of medium-skid silk / modified silk compositions and constituent AS compositions separated by Q-SEC (Q-flow-through). Figure 53A: Average molecular weights in kDa of medium-skid silk (MS) and AS101-AS105. Figure 53B: Polydispersity (PDI) measurements. Numerical data are presented in Table 27. [Figure 54] Figures 54A-B are analytical SEC-MALS of medium-skid silk / modified silk compositions and constituent AS compositions separated by SEC. Figure 54A: Average molecular weights in kDa of medium-skid silk (MS) and AS106-AS111. Figure 54B: Polydispersity (PDI) measurements. Numerical data are presented in Table 28. [Figure 55] FIG. 55 is a chart illustrating the average TEWL values ​​for Barrier Reduced Emulsions. [Figure 56] FIG. 56 is a chart illustrating the effect of Barrier Reducex Emulsion on wrinkles and photoaging. [Figure 57] Figure 57 is a chart illustrating the effect of Barrier Reducex Emulsion on hyperpigmentation. [Figure 58] Figure 58 is a chart illustrating the effect of Barrier Reducex emulsion on wrinkles. [Figure 59] FIG. 59 is a chart illustrating the effect of Barrier Reducex Emulsion on crow's feet wrinkles. [Figure 60] FIG. 60 is a chart illustrating the effect of Barrier Reducex emulsion on crow's feet lines. [Figure 61]Figure 61 is a chart illustrating the effect of Barrier Reducex Emulsion on marionette line wrinkles. [Figure 62] FIG. 62 is a chart illustrating the effect of Barrier Reducex emulsion on skin roughness. [Figure 63] FIG. 63 is a chart illustrating the effect of Barrier Reducex Emulsion on overall wrinkles. [Figure 64] FIG. 64 is a chart illustrating the effect of Barrier Reducex Emulsion on overall fine lines. [Figure 65] Figure 65 is a chart illustrating the effect of Barrier Redux emulsion on the appearance of media, deep streaks, and wrinkles. [Figure 66] Figure 66 is a chart illustrating the effect of Barrier Reducex emulsion on skin tone evenness. [Figure 67] Figure 67 is a chart illustrating the effect of Barrier Reducex emulsion on skin texture. [Figure 68] Figure 68 is a chart illustrating the effect of Barrier Redux emulsion on skin desquamation. [Figure 69A] Figure 69A shows the sequence listing for the fibroin heavy chain. [Figure 69B] Figure 69B shows the sequence listing for the fibroin heavy chain. [Figure 69C] Figure 69C shows the sequence listing for the fibroin heavy chain. [Figure 70] Figure 70 shows the sequence listing for the fibroin light chain. [Figure 71] Figure 71 shows the sequence listing for fibrohexamerin. [Figure 72] FIG. 72 is a graph illustrating the absorbency of different fabrics using laundry pods with activated silk. [Figure 73] FIG. 73 is a flow chart illustrating various embodiments for producing silk fibroin protein fragments (SPF) of the present disclosure. [Figure 74]Figure 74 is a flow chart illustrating various parameters that can be modified during the extraction and lysis steps during the process of producing a silk protein fragment solution of the present disclosure. [Figure 75] FIG. 75 is a chart showing the absorbency of Activated Silk™ with capryl / caprylyl glucoside coating on a wide range of interlock nylon fabrics; unfinished nylon interlock fabrics do not absorb water and have poor absorbency; after Activated Silk™ is coated with capryl / caprylyl glucoside, the absorbency of all nylon interlock fabrics increases significantly. [Figure 76] FIG. 76 is a chart showing the absorbency of Activated Silk™ with capryl / caprylyl glucoside coating on various nylon fabrics other than interlock structure, where unfinished nylon fabrics do not absorb water or have poor absorbency, and after Activated Silk™ is coated with capryl / caprylyl glucoside, the absorbency of all nylon fabrics increases significantly. [Figure 77] Figure 77 is a chart showing water absorbency curves without wash produced by varying the concentration of polyoxyethylene (29) castor oil in the emulsifier mixture (thereby varying the HLB) before adding it to the coating solution, with the silk concentration in the coating solution being 1 g / L for all samples. [Figure 78] Figure 78 is a chart showing no-wash hand ranking curves produced by varying the concentration of polyoxyethylene (29) castor oil (thereby varying the HLB) in the emulsifier mixture before adding it to the coating solution, with the silk concentration in the coating solution being 1 g / L for all samples. [Figure 79A]Figures 79A-79D are charts showing moisture control data for no wash (Figure 79A), 5 washes (Figure 79B), 10 washes (Figure 79C), and 25 washes (Figure 79D) produced by varying the concentration of emulsion mixture (polyoxyethylene (20) sorbitan monooleate, polyoxyethylene (20) sorbitan trioleate, polyoxyethylene (29) castor oil, and water in a 2:4:8:10 ratio) in the final coating solution, with the silk concentration in the coating solution being 1 g / L for all samples. [Figure 79B] As explained above in the section on Figure 79A. [Figure 79C] As explained above in the section on Figure 79A. [Figure 79D] As explained above in the section on Figure 79A. [Figure 80A] Figures 80A-80D are charts showing hand ranking results for no wash (Figure 80A), 5 washes (Figure 80B), 10 washes (Figure 80C), and 25 washes (Figure 80D) produced by varying the concentration of the emulsion mixture (polyoxyethylene (20) sorbitan monooleate, polyoxyethylene (20) sorbitan trioleate, polyoxyethylene (29) castor oil, and water in a 2:4:8:10 ratio) in the final coating solution, where 1 is the best hand ranking score and 8 is the worst hand ranking score. For all samples, the silk concentration in the solution is 1 g / L. [Figure 80B] As explained in the section on Figure 80A above. [Figure 80C] As explained in the section on Figure 80A above. [Figure 80D] As explained in the section on Figure 80A above. [Figure 81A]Figures 81A-81D are charts showing hand-feel ranking results from no wash (Figure 81A), 5 washes (Figure 81B), 10 washes (Figure 81C), and 25 washes (Figure 81D) produced by varying the concentration of medium molecular weight silk in the final coating solution, with 1 being the best ranking and 8 being the worst ranking. [Figure 81B] As explained above in the section on Figure 81A. [Figure 81C] As explained above in the section on Figure 81A. [Figure 81D] As explained above in the section on Figure 81A. [Figure 82A] Figures 82A-82D are charts showing moisture management results from no wash (Figure 82A), 5 washes (Figure 82B), 10 washes (Figure 82C), and 25 washes (Figure 82D) produced by varying the concentration of medium molecular weight silk in the final coating solution. [Figure 82B] As explained in the section on Figure 82A. [Figure 82C] As explained in the section on Figure 82A. [Figure 82D] As explained in the section on Figure 82A. [Figure 83A] Figures 83A-83D are graphs showing UV / Vis quantification experiments of textiles coated with low molecular weight activated silk and polyoxyethylene (20) monooleate solution. Figure 83A: Graph showing the percent of silk lost after five washes, relative to fiber surface area. Figure 83B: Graph showing the quantified mass of silk on the textile after coating, relative to fiber surface area. Figure 83C: Graph showing the percent of silk lost after five washes, relative to textile type. Figure 83D: Graph showing the quantified mass of silk for each textile before and after five washes, depending on fiber type. [Figure 83B] As explained above in the section on Figure 83A. [Figure 83C] As explained above in the section on Figure 83A. [Figure 83D] As explained above in the section on Figure 83A. [Figure 84] Figure 84 is a chart showing UV / Vis quantification experiments of fabrics coated with low molecular weight activated silk and polyoxyethylene (20) monooleate solution. The graph shows the quantified mass of silk on the fabric after coating in grams per square meter (GSM) relative to the mass of the fabric. This mass depends on the knit type, fiber content, and filament denier. [Figure 85A] Figures 85A-85C include a series of charts showing potentiometric titration curves with charge densities measured at pH 5 for an unfinished heavy-duty double-knit nylon fabric (Figure 85A), a heavy-duty double-knit nylon fabric with an activated silk finish (Figure 85B), and a heavy-duty double-knit nylon fabric with an Archroma RPU wetting agent finish (Figure 85C). Each fabric has titration curves obtained without washing (Figures 85A-85C, left panels) and with five washes (Figures 85A-85C, right panels). The change in charge density at pH 5 after washing is shown as ΔC. [Figure 85B] As explained in the section on Figure 85A above. [Figure 85C] As explained in the section on Figure 85A above. [Figure 86] Figure 86 illustrates the experimental setup for Example 18. Fabric samples were prepared and washed for five washes in a front-loading washing machine and the surface charge was measured as the difference in charge before and after washing. [Figure 87] Figure 87 is a schematic diagram showing the location and description of the zeta potential of a surface. [Figure 88] Figure 88 is a graph showing the change in zeta potential before and after five washes for five different fabric types and three different finish types. [Figure 89] Figure 89 is a schematic diagram showing the effective surface charge at ambient pH due to proton and hydroxide surface adsorption. [Figure 90] Figure 90 is a graph showing the change in charge density before and after five washes for five different fabric types at pH 5. The control section shows the unfinished fabric and the AS320 section shows the silk finished fabric. [Figure 91] FIG. 91 is a graph illustrating the UV / Vis spectrum of the synthetic silver silk nanoparticles tested in Example 19. [Figure 92] Figure 92 is a graph showing the absorbency of unfinished Night Blue DC98-IA pretreated with fixative only and AS-320 coated Night Blue DC98-IA pretreated with cationic fixative from T=0 to T=10. [Figure 93] Figure 93 is a graph showing the absorbency of unfinished black DC95-FA pretreated with fixative only, and black DC95-FA coated with AS-320 pretreated with cationic fixative from T=0 to T=10. [Figure 94] Figure 94 is a graph showing the absorbency of 17120513 black paper unfinished at T=0 and coated with AS-320 (15 g / L, with and without HT-232H fixative pretreatment) from T=0 to T=10. [Figure 95] Figure 95 is a graph showing the absorbance of 17120511 Blue unfinished at T=0 and coated with AS-320 (15 g / L, with and without HT-232H fixative pretreatment) from T=0 to T=10. [Figure 96] Figure 96 is a graph illustrating the improvement in Investigator Global Assessment (IGA) after using the body wash. [Figure 97] Figure 97 is a graph illustrating the improvement in redness after using the body wash. [Figure 98] Figure 98 is a graph illustrating the improvement in eczema counts after using the body wash. [Figure 99] FIG. 99 is a graph illustrating the improvement in individual corneometers after using body wash. [Figure 100] FIG. 100 is a graph illustrating individual TEWL improvements after using the body wash. [Figure 101]FIG. 101 is a graph illustrating the responses to a self-perception questionnaire after using the body wash. [Figure 102] FIG. 102 is a graph illustrating the responses to the self-perception questionnaire after using the body wash. [Figure 103] Figure 103 illustrates three chromatographic principles of silk fractionation. [Figure 104] Figure 104 illustrates size exclusion chromatography of silk fractions. [Figure 105] Figure 105 illustrates anion exchange chromatography followed by size exclusion chromatography of silk fractions. [Figure 106] Figure 106 illustrates anion exchange chromatography followed by hydrophobic interaction chromatography and size exclusion chromatography. [Figure 107] Figure 107 is a chart summarizing the three pipelines for silk fractionation. [Figure 108] Figure 108 shows a method for characterizing silk fractions. [Figure 109] Figure 109 illustrates size exclusion chromatography of low-skid silk. [Figure 110] FIG. 110 illustrates an example of characterization of silk compositions: size exclusion chromatography of low-skid silk. [Figure 111] Figure 111 illustrates an example of characterization of silk compositions: low-skid silk dynamic light scattering size exclusion chromatography. [Figure 112] Figure 112 is a chart containing the Z-average of low skid silk / modified polypeptide compositions. [Figure 113] Figure 113 illustrates the determination of molecular weight and polydispersity by HPLC. [Figure 114] Figure 114 illustrates silk fraction size exclusion chromatography of low-skid silk. The lower molecular weight fractions slowly self-assemble over time. [Figure 115] Figure 115 is a chart containing assays for characterizing silk fractions. [Figure 116] Figure 116 is a comparison of the molecular weight and polydispersity determinations by the two methods. [Figure 117] Figure 117 shows a graph presenting the data from Tables 70 and 71. Data is shown with standard deviation. "Nanoclay" refers to Elementis Bentone Hydroclay. [Figure 118] Figure 118 shows a schematic representation of a typical bentonite clay structure. [Figure 119] Figure 119 shows an SEM image of the cross section of a membrane cast with Elementis Bentone Hydroclay 2001. Highly ordered stacking of the clay layers is visible. [Figure 120] Figure 120 is an SEM image of the cross section of a membrane cast with pure RSF. [Figure 121] Figure 121 is an SEM image of the cross section of a 1:1 RSF / 2001 membrane cast under neutral (pH 7.0) conditions. The layered structure of the clay is preserved. [Figure 122] Figure 122 is an SEM image of the cross section of a 1:1 RSF / 2001 membrane cast under acidic (pH ∼3.5) conditions. Note the ribbon-like structure. [Figure 123] Figure 123 illustrates the increased diffusion paths created by the RSF / nanoclay composite. [Figure 124] Figure 124 shows FTIR scans of the amide I region of RSF / 2001 membranes cast under neutral conditions. The nanoclay concentration is varied from 0% (red) to 70% (yellow). As the nanoclay content increases, the amide I peak shifts to the left, away from the beta-sheet region. DETAILED DESCRIPTION OF THE INVENTION

[0024] In some embodiments, the compositions of the present disclosure comprise a peptide composition selected from compositions #1001 to #2450, having a weight average molecular weight selected from about 1 kDa to about 145 kDa, and having a polydispersity selected from 1 to about 5 (including, but not limited to, a polydispersity of 1), 1 to about 1.5 (including, but not limited to, a polydispersity of 1), about 1.5 to about 2, about 1.5 to about 3, about 2 to about 2.5, about 2.5 to about 3, about 3 to about 3.5, about 3.5 to about 4, about 4 to about 4.5, and about 4.5 to about 5.

[0025] Methods for producing silk fibroin or silk fibroin protein fragments and their applications in various fields are known and are described, for example, in U.S. Patent Nos. 9,187,538, 9,511,012, 9,517,191, 9,522,107, 9,522,108, 9,545,369, and 10,166,177, 10,287,728, and 10,301,768, all of which are incorporated herein in their entirety. Raw silk from the silkworm Bombyx mori is composed of two major proteins: silk fibroin (approximately 75%) and sericin (approximately 25%). Silk fibroin is a fibrous protein with a semi-crystalline structure that provides rigidity and strength. As used herein, the term "silk fibroin" refers to the fiber of Bombyx mori cocoons, which has a weight-average molecular weight of approximately 370,000 Da. Silkworm cocoon fibers consist of double strands of fibroin. The adhesive substance that holds these double strands together is sericin. Silk fibroin is composed of heavy chains (H chains) with a weight-average molecular weight of approximately 350,000 Da and light chains (L chains) with a weight-average molecular weight of approximately 25,000 Da. Silk fibroin is an amphiphilic polymer with large hydrophobic domains that account for the major component of the polymer, which has a high molecular weight. The hydrophobic regions are interrupted by small hydrophilic spacers, and the N- and C-termini of the chains are also highly hydrophilic. The hydrophobic domain of the H chain contains a repeating hexapeptide sequence of Gly-Ala-Gly-Ala-Gly-Ser and a repeat of the Gly-Ala / Ser / Tyr dipeptide, which can form stable antiparallel sheet crystallites. The amino acid sequence of the L chain is non-repetitive, making it more hydrophilic and relatively elastic. The hydrophilic (Tyr, Ser) and hydrophobic (Gly, Ala) chain segments in the silk fibroin molecule are alternately arranged to allow the silk fibroin molecule to self-assemble.

[0026] As used herein, the term "fibroin" includes silkworm fibroin and insect or spider silk proteins. In one embodiment, the fibroin is obtained from Bombyx mori. Raw silk from Bombyx mori is composed of two major proteins: silk fibroin (approximately 75%) and sericin (approximately 25%). As used herein, the term "silk fibroin" refers to the fiber of Bombyx mori cocoons, which has a weight-average molecular weight of approximately 370,000 Da. Methods for producing silk fibroin protein fragments and / or compositions thereof are known and are described, for example, in U.S. Patent Nos. 9,187,538, 9,511,012, 9,517,191, 9,522,107, 9,522,108, 9,545,369, and 10,166,177.

[0027] Recombinant silk is described in the patents and patent applications incorporated herein by reference: U.S. Patent Publication No. 2004 / 590196, U.S. Patent No. 7,754,851, U.S. Patent Publication No. 2007 / 654470, U.S. Patent No. 7,951,908, U.S. Patent Publication No. 2010 / 785960, U.S. Patent No. 8,034,897, U.S. Patent Publication Nos. 2009 / 0263430, 2008 / 226854, 2009 / 0123967, 2005 / 712095, 2007 / 991037, 2009 / 016 No. 2896, No. 2008 / 85266, U.S. Patent No. 8,372,436, U.S. Patent Publication Nos. 2007 / 989907, 2009 / 267596, 2010 / 319542, 2009 / 265344, 2012 / 684607, 2004 / 583227, U.S. Patent No. 8,030,024, U.S. Patent Publication No. 2006 / 643569, U.S. Patent No. 7,868,146, U.S. Patent Publication No. 2007 / 991916, U.S. Patent No. 8,097,583, U.S. Patent Publication No. 2006 / 643200, U.S. Patent Nos. 8,729,238, 8,877,903, U.S. Patent Publication Nos. 2019 / 0062557, 2016 / 0280960, 2011 / 0201783, 2008 / 991916, 2011 / 986662, 2012 / 697729, 2015 / 0328363, U.S. Patent No. 9,034,816, U.S. Patent Publication No. 2013 / 0172478, U.S. Patent No. 9,217,017, U.S. Patent Publication No. 2017 / 0202995, U.S. Patent No. 8,721,991, U.S. Patent Publication No. 2008 / 227498, U.S. Patent Nos. 9,233,067, 8,288,512, U.S. Patent Publication No. 2008 / 161364, U.S. Patent No. 7,148,039, U.S. Patent Publication Nos. 1999 / 247806, 2001 / 861597, 2004 / 887100, U.S. Patent Nos. 9,481,719, 8,765,688, U.S. Patent Publication Nos. 2008 / 80705, 2010 / 809102, U.S. Patent No. 8,367,803, U.S. Patent Publication No. 2010 / 664902, U.S. Patent No. 7,569,660, U.S. Patent Publication Nos. 1999 / 138833, 2000 / 591632, 2012 / 0065126, 2010 / 0278882, 2008 / 161352, 2010 / 0015070, 2009 / 513709, 2009 / 0194317, 2004 / 559286, 2005 / 89551, 2008 / 187824, 2005 / 0266242, 2005 / 0227322, and 20044418.

[0028] Recombinant silk is also described in other patents and patent applications, which are incorporated herein by reference: U.S. Patent Publication Nos. 2019 / 0062557, 2015 / 0284565, 2013 / 0225476, 2013 / 0172478, 2013 / 0136779, 2013 / 0109762, 2012 / 0252294, 2011 / 0230911, 2011 / 0201783, and 2010 / 029887. No. 7, U.S. Patent Nos. 10,478,520, 10,253,213, 10,072,152, 9,233,067, 9,217,017, 9,034,816, 8,877,903, 8,729,238, 8,721,991, 8,097,583, 8,034,897, 8,030,024, 7,951,908, 7,868,146, and 7,754,851.

[0029] The recombinant silk proteins and / or methods described herein may be any of those described above or in any of the U.S. Patent Nos. 8,173,772, 8,278,416, 8,618,255, 8,642,734, 8,691,581, 8,729,235, 9,115,204, 9,157,070, 9,309,299, 9,644,012, 9,708,376, 9,051,453, 9,617,315, 9,968,682, 9,689,089, 9,73 Nos. 2,125, 9,856,308, 9,926,348, 10,065,997, 10,316,069, and 10,329,332; and U.S. Patent Publication Nos. 2009 / 0226969, 2011 / 0281273, 2012 / 0041177, 2013 / 0065278, 2013 / 0115698, 2013 / 0316376, 2014 / 0058066, 2014 / 0079674, 2014 / 0245923, 2015 / 0087046, No. 2015 / 0119554, No. 2015 / 0141618, No. 2015 / 0291673, No. 2015 / 0291674 No. 2015 / 0239587, No. 2015 / 0344542, No. 2015 / 0361144, No. 2015 / 03748 No. 33, No. 2015 / 0376247, No. 2016 / 0024464, No. 2017 / 0066804, No. 2017 / 006 No. 6805, No. 2015 / 0293076, No. 2016 / 0222174, No. 2017 / 0283474, No. 2017 / 0 No. 088675, No. 2019 / 0135880, No. 2015 / 0329587, No. 2019 / 0040109, No. 2019 / 0135881, 2019 / 0177363, 2019 / 0225646, 2019 / 0233481, 20 19 / 0031842, 2018 / 0355120, 2019 / 0186050, 2019 / 0002644, 2020 / 0031887, 2018 / 0273590, 20191 / 094403, 2019 / 0031843,Same No. 2018 / 0251501, No. 2017 / 0066805, No. 2018 / 0127553, No. 2019 / 0329526, No. 2020 / 0031886, No. 201 8 / 0080147, 2019 / 0352349, 2020 / 0043085, 2019 / 0144819, 2019 / 0228449, 2019 / 034 0666, 2020 / 0000091, 2019 / 0194710, 2019 / 0151505, 2018 / 0265555, 2019 / 0352330, 2019 / 0248847, and 2019 / 0378191, which are incorporated herein by reference in their entireties.

[0030] As used herein, the term "substantially free of inorganic residue" means that a composition exhibits 0.1% (w / w) or less residue. In one embodiment, "substantially free of inorganic residue" refers to a composition exhibiting 0.05% (w / w) or less residue. In one embodiment, "substantially free of inorganic residue" refers to a composition exhibiting 0.01% (w / w) or less residue. In one embodiment, the amount of inorganic residue is from 0 ppm ("non-detectable" or "ND") to 1000 ppm. In one embodiment, the amount of inorganic residue is from ND to about 500 ppm. In one embodiment, the amount of inorganic residue is from ND to about 400 ppm. In one embodiment, the amount of inorganic residue is from ND to about 300 ppm. In one embodiment, the amount of inorganic residue is from ND to about 200 ppm. In one embodiment, the amount of inorganic residue is from ND to about 100 ppm. In one embodiment, the amount of inorganic residue is from 10 ppm to 1000 ppm.

[0031] As used herein, the term "substantially free of organic residue" means that a composition exhibits 0.1% (w / w) or less of residue; in one embodiment, "substantially free of organic residue" refers to a composition exhibiting 0.05% (w / w) or less of residue. In one embodiment, "substantially free of organic residue" refers to a composition exhibiting 0.01% (w / w) or less of residue. In one embodiment, the amount of organic residue is between 0 ppm ("non-detectable" or "ND") and 1000 ppm. In one embodiment, the amount of organic residue is between ND and about 500 ppm. In one embodiment, the amount of organic residue is between ND and about 400 ppm. In one embodiment, the amount of organic residue is between ND and about 300 ppm. In one embodiment, the amount of organic residue is between ND and about 200 ppm. In one embodiment, the amount of organic residue is between ND and about 100 ppm. In one embodiment, the amount of organic residue is between 10 ppm and 1000 ppm.

[0032] The compositions of the present disclosure are "biocompatible," meaning that the compositions are compatible with living tissues or systems by being non-toxic, non-intoxicating, or physiologically reactive, and by not causing immunological rejection. Such biocompatibility can be demonstrated by participants topically applying the compositions of the present disclosure to their skin for an extended period of time. In one embodiment, the extended period is about 3 days. In one embodiment, the extended period is about 7 days, in one embodiment, the extended period is about 14 days, and in one embodiment, the extended period is about 21 days. In one embodiment, the extended period is about 30 days. In one embodiment, the extended period is selected from the group consisting of about 1 month, about 2 months, about 3 months, about 4 months, about 5 months, about 6 months, about 7 months, about 8 months, about 9 months, about 10 months, about 11 months, about 12 months, and indefinitely.

[0033] The compositions of the present disclosure are "hypoallergenic," meaning that they are relatively unlikely to cause an allergic reaction. Such hypoallergenicity can be demonstrated by participants topically applying the compositions of the present disclosure to their skin for an extended period of time. In one embodiment, the extended period is about 3 days. In one embodiment, the extended period is about 7 days. In one embodiment, the extended period is about 14 days. In one embodiment, the extended period is about 21 days. In one embodiment, the extended period is about 30 days. In one embodiment, the extended period is selected from the group consisting of about 1 month, about 2 months, about 3 months, about 4 months, about 5 months, about 6 months, about 7 months, about 8 months, about 9 months, about 10 months, about 11 months, about 12 months, and indefinitely.

[0034] In one embodiment, the peptide compositions of the present disclosure have undetectable levels of LiBr residual. In one embodiment, the amount of LiBr residual in the compositions of the present disclosure is between 10 ppm and 1000 ppm. In one embodiment, the amount of LiBr residual in the compositions of the present disclosure is between 10 ppm and 300 ppm. In one embodiment, the amount of LiBr residual in the compositions of the present disclosure is less than 25 ppm. In one embodiment, the amount of LiBr residual in the compositions of the present disclosure is less than 50 ppm. In one embodiment, the amount of LiBr residual in the compositions of the present disclosure is less than 75 ppm. In one embodiment, the amount of LiBr residual in the compositions of the present disclosure is less than 100 ppm. In one embodiment, the amount of LiBr residual in the compositions of the present disclosure is less than 200 ppm. In one embodiment, the amount of LiBr residual in the compositions of the present disclosure is less than 300 ppm. In one embodiment, the amount of LiBr residual in the compositions of the present disclosure is less than 400 ppm. In one embodiment, the amount of LiBr residual in the compositions of this disclosure is less than 500 ppm. In one embodiment, the amount of LiBr residual in the compositions of this disclosure is less than 600 ppm. In one embodiment, the amount of LiBr residual in the compositions of this disclosure is less than 700 ppm. In one embodiment, the amount of LiBr residual in the compositions of this disclosure is less than 800 ppm. In one embodiment, the amount of LiBr residual in the compositions of this disclosure is less than 900 ppm. In one embodiment, the amount of LiBr residual in the compositions of this disclosure is less than 1000 ppm. In one embodiment, the amount of LiBr residual in the compositions of this disclosure is undetectable to 500 ppm. In one embodiment, the amount of LiBr residual in the compositions of this disclosure is undetectable to 450 ppm. In one embodiment, the amount of LiBr residual in the compositions of this disclosure is undetectable to 400 ppm. In one embodiment, the amount of LiBr residual in the compositions of this disclosure is undetectable to 350 ppm. In one embodiment, the amount of LiBr residual in the composition of the present disclosure is undetectable to 300 ppm. In one embodiment, the amount of LiBr residual in the composition of the present disclosure is undetectable to 250 ppm. In one embodiment, the amount of LiBr residual in the composition of the present disclosure is undetectable to 200 ppm. In one embodiment, the amount of LiBr residual in the composition of the present disclosure is undetectable to 150 ppm.In one embodiment, the amount of LiBr residual in the composition of the present disclosure is from undetectable to 100 ppm. In one embodiment, the amount of LiBr residual in the composition of the present disclosure is from 100 ppm to 200 ppm. In one embodiment, the amount of LiBr residual in the composition of the present disclosure is from 200 ppm to 300 ppm. In one embodiment, the amount of LiBr residual in the composition of the present disclosure is from 300 ppm to 400 ppm. In one embodiment, the amount of LiBr residual in the composition of the present disclosure is from 400 ppm to 500 ppm.

[0035] In one embodiment, the peptide compositions of the present disclosure have undetectable levels of Na2CO3 residue. In one embodiment, the amount of Na2CO3 residue in the compositions of the present disclosure is less than 100 ppm. In one embodiment, the amount of Na2CO3 residue in the compositions of the present disclosure is less than 200 ppm. In one embodiment, the amount of Na2CO3 residue in the compositions of the present disclosure is less than 300 ppm. In one embodiment, the amount of Na2CO3 residue in the compositions of the present disclosure is less than 400 ppm. In one embodiment, the amount of Na2CO3 residue in the compositions of the present disclosure is less than 500 ppm. In one embodiment, the amount of Na2CO3 residue in the compositions of the present disclosure is less than 600 ppm. In one embodiment, the amount of Na2CO3 residue in the compositions of the present disclosure is less than 700 ppm. In one embodiment, the amount of Na2CO3 residue in the compositions of the present disclosure is less than 800 ppm. In one embodiment, the amount of Na2CO3 residue in the compositions of the present disclosure is less than 900 ppm. In one embodiment, the amount of Na2CO3 residue in the composition of the present disclosure is less than 1000 ppm. In one embodiment, the amount of Na2CO3 residue in the composition of the present disclosure is undetectable to 500 ppm. In one embodiment, the amount of Na2CO3 residue in the composition of the present disclosure is undetectable to 450 ppm. In one embodiment, the amount of Na2CO3 residue in the composition of the present disclosure is undetectable to 400 ppm. In one embodiment, the amount of Na2CO3 residue in the composition of the present disclosure is undetectable to 350 ppm. In one embodiment, the amount of Na2CO3 residue in the composition of the present disclosure is undetectable to 300 ppm. In one embodiment, the amount of Na2CO3 residue in the composition of the present disclosure is undetectable to 250 ppm. In one embodiment, the amount of Na2CO3 residue in the composition of the present disclosure is undetectable to 200 ppm. In one embodiment, the amount of Na2CO3 residue in the composition of the present disclosure is undetectable to 150 ppm. In one embodiment, the amount of Na2CO3 residue in the composition of the present disclosure is undetectable to 100 ppm. In one embodiment, the amount of Na2CO3 residue in the composition of the present disclosure is 100 ppm to 200 ppm. In one embodiment, the amount of Na2CO3 residue in the composition of the present disclosure is 200 ppm to 300 ppm. In one embodiment, the amount of Na2CO3 residue in the composition of the present disclosure is 300 ppm to 400 ppm.In one embodiment, the amount of Na2CO3 residue in the composition of the present disclosure is from 400 ppm to 500 ppm.

[0036] As used herein, when referring to a numerical value or numerical range, the term "about" means that the stated number or numerical range is included with the number or numerical range within experimental variation, or within statistical experimental error from the stated number or numerical range, and the variation or error is 0% to 15%, or 0% to 10%, or 0% to 5% of the stated number or numerical range.

[0037] Silk proteins or fragments thereof, silk solutions or mixtures (e.g., SPF or SFS solutions or mixtures), and the like, may be prepared according to the methods described in U.S. Pat. Nos. 9,187,538, 9,522,107, 9,522,108, 9,511,012, 9,517,191, 9,545,369, and 10,166,177, as well as U.S. Patent Publication Nos. 2016 / 0222579 and 2016 / 0281294, and International Patent Publication Nos. WO 2016 / 090055 and 2017 / 011679, which are incorporated herein by reference in their entireties. The use of silk fibroin or silk fibroin fragments in coating applications is known and is described, for example, in US Pat. Nos. 10,287,728 and 10,301,768.

[0038] The present disclosure provides articles comprising coated textiles, wherein the coating comprises a surfactant and / or emulsifier system and modified silk fragments as disclosed herein, e.g., silk fibroin fragments, including, but not limited to, fibroin peptides and / or protein fragments in which at least one of the amino acids has been modified, substituted, or replaced as disclosed herein, as well as methods of making such articles.

[0039] Silk is a natural polymer produced by various insects and spiders. Silk produced by Bombyx mori (the silkworm) contains a filament core protein, silk fibroin, and a colloidal coating composed of the nonfilamentous protein sericin. Silk fibroin is an FDA-approved, edible, nontoxic, and relatively inexpensive protein derived from silkworm cocoons. The amino acid structure and content of silk fibroin protein are highly similar to those found in human body tissues.

[0040] Methods for producing silk fibroin protein fragments are known and are described, for example, in U.S. Patent Nos. 9,187,538, 9,511,012, 9,517,191, 9,522,107, 9,522,108, 9,545,369, and 10,166,177. Methods for using silk fibroin or silk fibroin fragments in coating applications, including animal hair coating applications, are known and are described, for example, in U.S. Patent Application Publication Nos. 2016 / 0222579 and 2016 / 0281294. Compositions and methods for using silk fibroin or silk fibroin fragments in cosmetic applications are known and are described, for example, in U.S. Patent Application Publication Nos. 2018 / 0280274 and 2018 / 0008522, and International Patent Application Publication No. WO 2019 / 005848. All publications cited herein are incorporated by reference in their entirety.

[0041] Recent advances in silk material technology include the emergence of top-down and bottom-up engineering of silk cocoons, specifically regenerating them into aqueous silk solutions, and using genetic engineering to produce recombinant silk with molecularly defined compositions (Tran et al., A review of the emerging role of silk for the treatment of the eye, Pharm. Res., 2018, vol. 35, pp. 1-16). Recently, silk fibroin protein has been reported to find applications in ocular tissue reconstruction, corneal tissue engineering, and ocular surface repair due to its biocompatibility, tunable properties, and transparency. Silk membranes have been found to support corneal cell growth and express stratified epithelial cell sheets comparable to amniotic membrane stroma (Lawrence et al., Silk film biomaterials for corneal tissue engineering, Biomaterials, 2009;30(7):1299-308; Harkin et al., Silk fibroin in ocular tissue reconstruction, Biomaterials, 2011; Chirtla et al., Bombyx mori silk fibroin membranes as potential substrata for epithelial constructs used in the management of ocular surface disorders. Tissue Engineering Pan A, 2008;14(7):1203-11.).Silk fibroin protein and hydrolyzed peptide fragments have been shown to inhibit the transcription and upstream activation of NF-kB protein subunits and inflammatory molecules classically under the control of NF-kB (Hayden et al., Cell Research, 2011. 21(2):223-244; Chon et al., International Journal of Molecular Medicine, 2012. 30(5):1203-1210; Kim et al., J. Neurosurg., 2011. 114(2):485-90; J. Microbiol. Biotechnol., 2012. 22(4):494-500).

[0042] definition As used in the above section, and throughout the remainder of the specification, unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this disclosure belongs. All patents and publications mentioned herein are incorporated by reference in their entirety.

[0043] Unless otherwise specified, all percentages, parts, and ratios are based on the total weight of the eye care compositions of the present disclosure. All such weights relating to listed ingredients are based on the active level and therefore do not include solvents or by-products that may be included in commercially available materials, unless otherwise specified. The term "weight percent" may be referred to herein as "wt.%" or % w / w.

[0044] As used herein, the terms "a," "an," or "the" are generally interpreted to encompass both the singular and the plural.

[0045] As used herein, the term "about" generally refers to a particular numerical value that is within an acceptable error range as determined by one of ordinary skill in the art, which depends in part on how the numerical value is measured or determined, i.e., the limitations of the measurement system. For example, "about" can mean zero variation and a range of ±20%, ±10%, or ±5% of the given numerical value.

[0046] As used herein, the term "dermatologically acceptable carrier" means a carrier that is suitable for use in contact with mammalian keratinous tissue without causing any adverse effects, such as, for example, undue toxicity, incompatibility, instability, allergic reaction, etc. Dermatologically acceptable carriers may include, but are not limited to, water, liquid or solid emollients, moisturizers, solvents, and the like.

[0047] As used herein, the term "hydrophilic-lipophilic balance" (HLB) of a surfactant and / or emulsifier is a measure of the degree of hydrophilicity or hydrophobicity, determined by calculating values ​​for different regions of the molecule, according to the Griffin method: HLB = 20 x M h / M, where M h where M is the molecular mass of the hydrophilic portion of the surfactant and / or emulsifier, and M is the molecular mass of the entire surfactant and / or emulsifier molecule, giving results on a scale of 0 to 20. An HLB value of 0 corresponds to a completely lipophilic molecule, and a value of 20 corresponds to a completely hydrophilic molecule. The HLB value can be used to predict the surfactant and / or emulsifier properties of a molecule as follows: HLB < 10: lipid soluble (water insoluble), HLB > 10: water soluble (lipid insoluble), HLB = 1-3: antifoaming agent, 3-6: W / O (water-in-oil) emulsifier, 7-9: wetting agent and spreading agent, 8-16: O / W (oil-in-water) emulsifier, 13-16: surfactant, 16-18: solubilizer or hydrotrope.

[0048] As used herein, "average weight average molecular weight" refers to the average of two or more values ​​of weight average molecular weight of silk fibroin or fragments thereof of the same composition, the two or more values ​​being determined by two or more separate experimental readings.

[0049] As used herein, the term "polydispersity (PD)" of a polymer is commonly used as a measure of the broadness of the molecular weight distribution of a polymer and is defined by the following formula:

number

[0050] As used herein, the term "substantially homogeneous" can refer to silk fibroin-based protein fragments that are distributed in a normal distribution around a specified molecular weight. As used herein, the term "substantially homogeneous" can refer to, for example, an even distribution of components or additives, such as silk fibroin fragments, dermatologically acceptable carriers, etc., throughout the compositions of the present disclosure.

[0051] As used herein, the terms "silk fibroin peptide," "silk fibroin protein fragment," and "silk fibroin fragment" are used interchangeably. Molecular weight or number of amino acid units is defined when molecular size is the key parameter.

[0052] As used herein, the term "fast dissolving solid form" refers to fast dissolving solid forms including lyophilized forms (cakes, wafers, films) and compressed tablets.

[0053] As used herein, the term "peptide" or "protein" refers to a chain of amino acids held together by peptide bonds (also called amide bonds). The fundamental distinguishing factors between proteins and peptides are size and structure. Peptides are smaller than proteins. Conventionally, peptides are defined as molecules consisting of 2 to 50 amino acids, while proteins are composed of 50 or more amino acids. In addition, peptides tend to be less well-defined in structure than proteins and can adopt complex conformations known as secondary, tertiary, and quaternary structures.

[0054] As used herein, the term "fibroin" or "silk protein" refers to a type of structural protein produced by certain silk-producing spider and insect species (see definitions provided in the WIPO Pearl-WIPO's Multilingual Terminology Portal database, https: / / wipopearl.wipo.int / en / linguistic). Fibroin may include silkworm fibroin, insect or spider silk proteins (e.g., spidroins), recombinant spider proteins, silk proteins present in other spider silk types, such as tubular gland silk proteins (TuSPs), flagellate gland silk proteins, ampullate gland silk proteins, grape-like gland silk proteins, piriform gland silk proteins, agglutinating gland silk glue, silkworm fibroin produced by genetically modified silkworms, or recombinant silkworm fibroin.

[0055] As used herein, the term "silk fibroin" refers to silkworm fibroin, silk fibroin produced by genetically modified silkworms, or recombinant silkworm fibroin (see (1) Narayan Ed., Encyclopedia of Biomedical Engineering, Vol. 2, Elsevier, 2019; (2) Kobayashi et al. Eds., Encyclopedia of Polymeric Nanomaterials, Springer, 2014, https: / / link.springer.com / referenceworkentry / 10.1007%2F978-3-642-36199-9_323-1). In one embodiment, the silk fibroin is obtained from Bombyx mori.

[0056] As used herein, the term "solid solution" refers to an active agent molecularly dissolved in a solid excipient matrix, such as a hydrophobic polymer, wherein the active agent is miscible with the polymer matrix excipient.

[0057] As used herein, the term "solid dispersion" refers to an active agent dispersed as crystalline or amorphous particles, wherein the active agent is dispersed in an amorphous polymer and randomly distributed among the polymer matrix excipients.

[0058] As used herein, the term "substantially homogeneous" may refer to silk fibroin-based protein fragments that are distributed in a normal distribution around a specified molecular weight. As used herein, the term "substantially homogeneous" may also refer to, for example, an even distribution of components or additives, such as silk fibroin-based protein fragments, dermatologically acceptable carriers, etc., throughout the silk eye care composition.

[0059] As used herein, the term "surface tension" refers to the tendency of a fluid surface to contract to the smallest possible surface area. At a liquid-air interface, surface tension results from the greater attraction of liquid molecules to each other than to molecules in the air (due to adhesive forces). The net effect is an inward force at the liquid's surface that causes it to behave as if it were covered by a stretched elastic film. Because of the relatively high attraction of water molecules to each other through a network of hydrogen bonds, water has a higher surface tension (72.8 mN / m at 20°C) than most other liquids.

[0060] SPF Definition and Characteristics As used herein, "silk protein fragments" (SPFs) include, but are not limited to, one or more of the following: "silk fibroin fragments" as defined herein, "recombinant silk fragments" as defined herein, "spider silk fragments" as defined herein, "silk fibroin-like protein fragments" as defined herein, "chemically modified silk fragments" as defined herein, "sericin or sericin fragments" as defined herein, and / or modified silk fragments disclosed herein, such as, but not limited to, fibroin peptides and / or protein fragments in which at least one of the amino acids has been modified, substituted, or replaced as disclosed herein. SPFs may have any molecular weight value or range described herein, and any polydispersity value or range described herein.

[0061] SPF molecular weight and polydispersity In one embodiment, the composition of the present disclosure comprises an SPF having an average weight average molecular weight selected from about 1 to about 5 kDa. In one embodiment, the composition of the present disclosure comprises an SPF having an average weight average molecular weight selected from about 5 to about 10 kDa. In one embodiment, the composition of the present disclosure comprises an SPF having an average weight average molecular weight selected from about 10 to about 15 kDa. In one embodiment, the composition of the present disclosure comprises an SPF having an average weight average molecular weight selected from about 15 to about 20 kDa. In one embodiment, the composition of the present disclosure comprises an SPF having an average weight average molecular weight selected from about 14 to about 30 kDa. In one embodiment, the composition of the present disclosure comprises an SPF having an average weight average molecular weight selected from about 20 to about 25 kDa. In one embodiment, the composition of the present disclosure comprises an SPF having an average weight average molecular weight selected from about 25 to about 30 kDa. In one embodiment, the composition of the present disclosure comprises an SPF having an average weight average molecular weight selected from about 30 to about 35 kDa. In one embodiment, the composition of the present disclosure comprises an SPF having an average weight average molecular weight selected from about 35 to about 40 kDa. In one embodiment, the composition of the present disclosure comprises an SPF having an average weight average molecular weight selected from about 39 to about 54 kDa. In one embodiment, the composition of the present disclosure comprises an SPF having an average weight average molecular weight selected from about 40 to about 45 kDa. In one embodiment, the composition of the present disclosure comprises an SPF having an average weight average molecular weight selected from about 45 to about 50 kDa. In one embodiment, the composition of the present disclosure comprises an SPF having an average weight average molecular weight selected from about 50 to about 55 kDa. In one embodiment, the composition of the present disclosure comprises an SPF having an average weight average molecular weight selected from about 55 to about 60 kDa. In one embodiment, the composition of the present disclosure comprises an SPF having an average weight average molecular weight selected from about 60 to about 65 kDa. In one embodiment, the composition of the present disclosure comprises an SPF having an average weight average molecular weight selected from about 65 to about 70 kDa. In one embodiment, the composition of the present disclosure comprises an SPF having an average weight average molecular weight selected from about 70 to about 75 kDa. In one embodiment, the composition of the present disclosure comprises an SPF having an average weight average molecular weight selected from about 75 to about 80 kDa.In one embodiment, the composition of the present disclosure comprises an SPF having an average weight average molecular weight selected from about 80 to about 85 kDa. In one embodiment, the composition of the present disclosure comprises an SPF having an average weight average molecular weight selected from about 85 to about 90 kDa. In one embodiment, the composition of the present disclosure comprises an SPF having an average weight average molecular weight selected from about 90 to about 95 kDa. In one embodiment, the composition of the present disclosure comprises an SPF having an average weight average molecular weight selected from about 95 to about 100 kDa. In one embodiment, the composition of the present disclosure comprises an SPF having an average weight average molecular weight selected from about 100 to about 105 kDa. In one embodiment, the composition of the present disclosure comprises an SPF having an average weight average molecular weight selected from about 105 to about 110 kDa. In one embodiment, the composition of the present disclosure comprises an SPF having an average weight average molecular weight selected from about 110 to about 115 kDa. In one embodiment, the composition of the present disclosure comprises an SPF having an average weight average molecular weight selected from about 115 to about 120 kDa. In one embodiment, the composition of the present disclosure comprises an SPF having an average weight average molecular weight selected from about 120 to about 125 kDa. In one embodiment, the composition of the present disclosure comprises an SPF having an average weight average molecular weight selected from about 125 to about 130 kDa. In one embodiment, the composition of the present disclosure comprises an SPF having an average weight average molecular weight selected from about 130 to about 135 kDa. In one embodiment, the composition of the present disclosure comprises an SPF having an average weight average molecular weight selected from about 135 to about 140 kDa. In one embodiment, the composition of the present disclosure comprises an SPF having an average weight average molecular weight selected from about 140 to about 145 kDa. In one embodiment, the composition of the present disclosure comprises an SPF having an average weight average molecular weight selected from about 145 to about 150 kDa. In one embodiment, the composition of the present disclosure comprises an SPF having an average weight average molecular weight selected from about 150 to about 155 kDa. In one embodiment, the composition of the present disclosure comprises an SPF having an average weight average molecular weight selected from about 155 to about 160 kDa. In one embodiment, the composition of the present disclosure comprises an SPF having an average weight average molecular weight selected from about 160 to about 165 kDa. In one embodiment, the composition of the present disclosure comprises an SPF having an average weight average molecular weight selected from about 165 to about 170 kDa.In one embodiment, the composition of the present disclosure comprises an SPF having an average weight average molecular weight selected from about 170 to about 175 kDa. In one embodiment, the composition of the present disclosure comprises an SPF having an average weight average molecular weight selected from about 175 to about 180 kDa.

[0062] In one embodiment, the composition of the present disclosure comprises an SPF having an average weight average molecular weight selected from about 180 to about 185 kDa. In one embodiment, the composition of the present disclosure comprises an SPF having an average weight average molecular weight selected from about 185 to about 190 kDa. In one embodiment, the composition of the present disclosure comprises an SPF having an average weight average molecular weight selected from about 190 to about 195 kDa. In one embodiment, the composition of the present disclosure comprises an SPF having an average weight average molecular weight selected from about 195 to about 200 kDa. In one embodiment, the composition of the present disclosure comprises an SPF having an average weight average molecular weight selected from about 200 to about 205 kDa. In one embodiment, the composition of the present disclosure comprises an SPF having an average weight average molecular weight selected from about 205 to about 210 kDa. In one embodiment, the composition of the present disclosure comprises an SPF having an average weight average molecular weight selected from about 210 to about 215 kDa. In one embodiment, the composition of the present disclosure comprises an SPF having an average weight average molecular weight selected from about 215 to about 220 kDa.

[0063] In one embodiment, the composition of the present disclosure comprises an SPF having an average weight average molecular weight selected from about 220 to about 225 kDa. In one embodiment, the composition of the present disclosure comprises an SPF having an average weight average molecular weight selected from about 225 to about 230 kDa. In one embodiment, the composition of the present disclosure comprises an SPF having an average weight average molecular weight selected from about 230 to about 235 kDa. In one embodiment, the composition of the present disclosure comprises an SPF having an average weight average molecular weight selected from about 235 to about 240 kDa. In one embodiment, the composition of the present disclosure comprises an SPF having an average weight average molecular weight selected from about 240 to about 245 kDa. In one embodiment, the composition of the present disclosure comprises an SPF having an average weight average molecular weight selected from about 245 to about 250 kDa. In one embodiment, the composition of the present disclosure comprises an SPF having an average weight average molecular weight selected from about 250 to about 255 kDa. In one embodiment, the composition of the present disclosure comprises an SPF having an average weight average molecular weight selected from about 255 to about 260 kDa.

[0064] In one embodiment, the composition of the present disclosure comprises an SPF having an average weight average molecular weight selected from about 260 to about 265 kDa. In one embodiment, the composition of the present disclosure comprises an SPF having an average weight average molecular weight selected from about 265 to about 270 kDa. In one embodiment, the composition of the present disclosure comprises an SPF having an average weight average molecular weight selected from about 270 to about 275 kDa. In one embodiment, the composition of the present disclosure comprises an SPF having an average weight average molecular weight selected from about 275 to about 280 kDa. In one embodiment, the composition of the present disclosure comprises an SPF having an average weight average molecular weight selected from about 280 to about 285 kDa. In one embodiment, the composition of the present disclosure comprises an SPF having an average weight average molecular weight selected from about 285 to about 290 kDa. In one embodiment, the composition of the present disclosure comprises an SPF having an average weight average molecular weight selected from about 290 to about 295 kDa. In one embodiment, the composition of the present disclosure comprises an SPF having an average weight average molecular weight selected from about 295 to about 300 kDa.

[0065] In one embodiment, the composition of the present disclosure comprises an SPF having an average weight average molecular weight selected from about 300 to about 305 kDa. In one embodiment, the composition of the present disclosure comprises an SPF having an average weight average molecular weight selected from about 305 to about 310 kDa. In one embodiment, the composition of the present disclosure comprises an SPF having an average weight average molecular weight selected from about 310 to about 315 kDa. In one embodiment, the composition of the present disclosure comprises an SPF having an average weight average molecular weight selected from about 315 to about 320 kDa. In one embodiment, the composition of the present disclosure comprises an SPF having an average weight average molecular weight selected from about 320 to about 325 kDa. In one embodiment, the composition of the present disclosure comprises an SPF having an average weight average molecular weight selected from about 325 to about 330 kDa. In one embodiment, the composition of the present disclosure comprises an SPF having an average weight average molecular weight selected from about 330 to about 335 kDa. In one embodiment, the composition of the present disclosure comprises an SPF having an average weight average molecular weight selected from about 335 to about 340 kDa.

[0066] In one embodiment, the composition of the present disclosure comprises an SPF having an average weight average molecular weight selected from about 340 to about 345 kDa. In one embodiment, the composition of the present disclosure comprises an SPF having an average weight average molecular weight selected from about 345 to about 350 kDa.

[0067] In some embodiments, the compositions of the present disclosure include an SPF composition selected from compositions #1001-#2450, having a weight average molecular weight selected from about 1 kDa to about 145 kDa, and having a polydispersity selected from 1 to about 5 (including, but not limited to, a polydispersity of 1), 1 to about 1.5 (including, but not limited to, a polydispersity of 1), about 1.5 to about 2, about 1.5 to about 3, about 2 to about 2.5, about 2.5 to about 3, about 3 to about 3.5, about 3.5 to about 4, about 4 to about 4.5, and about 4.5 to about 5: JPEG2025532044000003.jpg75170 JPEG2025532044000004.jpg255170 JPEG2025532044000005.jpg255170 JPEG2025532044000006.jpg198170

[0068] In one embodiment, the SPF in the composition of the present disclosure has a polydispersity selected from 1 to about 5.0, including, but not limited to, a polydispersity of 1. In one embodiment, the SPF in the composition of the present disclosure has a polydispersity selected from about 1.5 to about 3.0. In one embodiment, the SPF in the composition of the present disclosure has a polydispersity selected from 1 to about 1.5, including, but not limited to, a polydispersity of 1. In one embodiment, the SPF in the composition of the present disclosure has a polydispersity selected from about 1.5 to about 2.0. In one embodiment, the SPF in the composition of the present disclosure has a polydispersity selected from about 2.0 to about 2.5. In one embodiment, the SPF in the composition of the present disclosure has a polydispersity selected from about 2.5 to about 3.0.

[0069] In one embodiment, the SPF in the composition of the present disclosure has a polydispersity selected from about 3.0 to about 3.5. In one embodiment, the SPF in the composition of the present disclosure has a polydispersity selected from about 3.5 to about 4.0. In one embodiment, the SPF in the composition of the present disclosure has a polydispersity selected from about 4.0 to about 4.5. In one embodiment, the SPF in the composition of the present disclosure has a polydispersity selected from about 4.5 to about 5.0.

[0070] In one embodiment, the SPF in the composition of the present disclosure has a polydispersity of 1. In one embodiment, the SPF in the composition of the present disclosure has a polydispersity of about 1.1. In one embodiment, the SPF in the composition of the present disclosure has a polydispersity of about 1.2. In one embodiment, the SPF in the composition of the present disclosure has a polydispersity of about 1.3. In one embodiment, the SPF in the composition of the present disclosure has a polydispersity of about 1.4. In one embodiment, the SPF in the composition of the present disclosure has a polydispersity of about 1.5. In one embodiment, the SPF in the composition of the present disclosure has a polydispersity of about 1.6. In one embodiment, the SPF in the composition of the present disclosure has a polydispersity of about 1.7. In one embodiment, the SPF in the composition of the present disclosure has a polydispersity of about 1.8. In one embodiment, the SPF in the composition of the present disclosure has a polydispersity of about 1.9. In one embodiment, the SPF in the composition of the present disclosure has a polydispersity of about 2.0. In one embodiment, the SPF in the composition of the present disclosure has a polydispersity of about 2.1. In one embodiment, the SPF in the composition of the present disclosure has a polydispersity of about 2.2. In one embodiment, the SPF in the composition of the present disclosure has a polydispersity of about 2.3. In one embodiment, the SPF in the composition of the present disclosure has a polydispersity of about 2.4. In one embodiment, the SPF in the composition of the present disclosure has a polydispersity of about 2.5. In one embodiment, the SPF in the composition of the present disclosure has a polydispersity of about 2.6. In one embodiment, the SPF in the composition of the present disclosure has a polydispersity of about 2.7. In one embodiment, the SPF in the composition of the present disclosure has a polydispersity of about 2.8. In one embodiment, the SPF in the composition of the present disclosure has a polydispersity of about 2.9. In one embodiment, the SPF in the composition of the present disclosure has a polydispersity of about 3.0. In one embodiment, the SPF in the composition of the present disclosure has a polydispersity of about 3.1. In one embodiment, the SPF in the composition of the present disclosure has a polydispersity of about 3.2. In one embodiment, the SPF in the composition of the present disclosure has a polydispersity of about 3.3. In one embodiment, the SPF in the composition of the present disclosure has a polydispersity of about 3.4. In one embodiment, the SPF in the composition of the present disclosure has a polydispersity of about 3.5. In one embodiment, the SPF in the composition of the present disclosure has a polydispersity of about 3.6.In one embodiment, the SPF in the composition of the present disclosure has a polydispersity of about 3.7. In one embodiment, the SPF in the composition of the present disclosure has a polydispersity of about 3.8. In one embodiment, the SPF in the composition of the present disclosure has a polydispersity of about 3.9. In one embodiment, the SPF in the composition of the present disclosure has a polydispersity of about 4.0. In one embodiment, the SPF in the composition of the present disclosure has a polydispersity of about 4.1. In one embodiment, the SPF in the composition of the present disclosure has a polydispersity of about 4.2. In one embodiment, the SPF in the composition of the present disclosure has a polydispersity of about 4.3. In one embodiment, the SPF in the composition of the present disclosure has a polydispersity of about 4.4. In one embodiment, the SPF in the composition of the present disclosure has a polydispersity of about 4.5. In one embodiment, the SPF in the composition of the present disclosure has a polydispersity of about 4.6. In one embodiment, the SPF in the composition of the present disclosure has a polydispersity of about 4.7. In one embodiment, the SPF in the composition of the present disclosure has a polydispersity of about 4.8. In one embodiment, the SPF in the composition of the present disclosure has a polydispersity of about 4.9. In one embodiment, the SPF in the composition of the present disclosure has a polydispersity of about 5.0.

[0071] Silk fibroin fragments Methods for producing silk fibroin or silk fibroin protein fragments and their applications in various fields are known and are described, for example, in U.S. Patent Nos. 9,187,538, 9,511,012, 9,517,191, 9,522,107, 9,522,108, 9,545,369, and 10,166,177, 10,287,728, and 10,301,768, all of which are incorporated herein in their entirety. Raw silk from the silkworm Bombyx mori is composed of two major proteins: silk fibroin (approximately 75%) and sericin (approximately 25%). Silk fibroin is a fibrous protein with a semi-crystalline structure that provides rigidity and strength. As used herein, the term "silk fibroin" refers to the fiber of Bombyx mori cocoons, which has a weight-average molecular weight of approximately 370,000 Da. Silkworm cocoon fibers consist of double strands of fibroin. The adhesive substance that holds these double strands together is sericin. Silk fibroin is composed of heavy chains (H chains) with a weight-average molecular weight of approximately 350,000 Da and light chains (L chains) with a weight-average molecular weight of approximately 25,000 Da. Silk fibroin is an amphiphilic polymer with large hydrophobic domains that account for the major component of the polymer, which has a high molecular weight. The hydrophobic regions are interrupted by small hydrophilic spacers, and the N- and C-termini of the chains are also highly hydrophilic. The hydrophobic domain of the H chain contains a repeating hexapeptide sequence of Gly-Ala-Gly-Ala-Gly-Ser and a repeat of the Gly-Ala / Ser / Tyr dipeptide, which can form stable antiparallel sheet crystallites. The amino acid sequence of the L chain is non-repetitive, making it more hydrophilic and relatively elastic. The hydrophilic (Tyr, Ser) and hydrophobic (Gly, Ala) chain segments in the silk fibroin molecule are alternately arranged to allow the silk fibroin molecule to self-assemble.

[0072] Provided herein are methods for producing pure and highly scalable silk fibroin-protein fragment mixture solutions that may be used across multiple industries for a variety of applications. Without wishing to be bound by any particular theory, it is believed that these methods are equally applicable to the fragmentation of any SPF described herein, including, but not limited to, recombinant silk proteins and silk-like or fibroin-like proteins.

[0073] As used herein, the term "fibroin" includes silkworm fibroin and insect or spider silk proteins. In one embodiment, the fibroin is obtained from Bombyx mori. Raw silk from Bombyx mori is composed of two major proteins: silk fibroin (approximately 75%) and sericin (approximately 25%). Silk fibroin is a fibrous protein with a semi-crystalline structure that provides rigidity and strength. As used herein, the term "silk fibroin" refers to Bombyx mori cocoon fibers having a weight-average molecular weight of approximately 370,000 Da. Conversion of these insoluble silk fibroin fibrils into water-soluble silk fibroin protein fragments requires the addition of concentrated neutral salt (e.g., 8-10 M lithium bromide), which interferes with the intermolecular and intramolecular ionic and hydrogen bonds that would otherwise render the fibroin protein insoluble in water. Methods for producing silk fibroin protein fragments, and / or compositions thereof, are known and are described, for example, in U.S. Patent Nos. 9,187,538, 9,511,012, 9,517,191, 9,522,107, 9,522,108, 9,545,369, and 10,166,177.

[0074] Figure 73 is a flow chart illustrating a general embodiment for producing some embodiments of the present disclosure. Of course, not all of the illustrated steps are necessarily required to make all of the silk solutions of the present disclosure. As illustrated in step A of Figure 74, cocoons (heat-treated or not), silk fibers, silk powder, spider silk, or recombinant spider silk can be used as the silk source.

[0075] FIG. 74 is a flow chart illustrating a general embodiment for various parameters that can be modified during the extraction and lysis steps during the process of producing a silk protein fragment solution of the present disclosure.

[0076] In some embodiments, it is specifically disclosed herein that the molecular weight of the silk fibroin protein fragment composition was determined using high-pressure liquid chromatography (HPLC) with a refractive index detector (RID), and the polydispersity was calculated using Cirrus GPC Online GPC / SEC software version 3.3 (Agilent). In some embodiments, it is specifically disclosed herein that the molecular weight of the silk fibroin protein fragment composition was determined using the SEC-MALS method.

[0077] Data Analysis and Calculations - Calculation of Average Molecular Weight using Cirrus Software Chromatographic data files of standards and analytical samples are uploaded to the Cirrus SEC data collection and molecular weight analysis software. For each injection of sample, the weight average molecular weight (M w ), number average molecular weight (M n ), peak average molecular weight (M p ), and polydispersity are calculated.

[0078] Spider silk fragments Spider silk is a natural polymer composed of three domains: a repetitive middle core domain that dominates the protein chain, and nonrepetitive N- and C-terminal domains. The large core domain is organized in a block copolymer-like arrangement, in which two basic sequences, crystalline [poly(A) or poly(GA)] and less crystalline (GGX or GPGXX (SEQ ID NO: 7)) polypeptides, alternate. Dragline silk is a protein complex composed of major ampullate gland dragline silk protein 1 (MaSp1) and major ampullate gland dragline silk protein 2 (MaSp2). Both silks are approximately 3500 amino acids long. MaSp1 can be found in the fiber core and periphery, while MaSp2 clusters within a specific core area. The large central domains of MaSp1 and MaSp2 are organized in a block copolymer-like arrangement, in which two basic sequences, crystalline [poly(A) or poly(GA)] and less crystalline (GGX or GPGXX (SEQ ID NO: 7)) polypeptides, alternate within the core domain. Specific secondary structures have been assigned to the poly(A) / (GA), GGX, and GPGXX (SEQ ID NO: 7) motifs, including β-sheets, β-helices, and β-spirals, respectively. The primary sequence, composition, and secondary structural elements of the repetitive core domain are responsible for the mechanical properties of spider silk; whereas, the nonrepetitive N- and C-terminal domains are essential for the storage of liquid silk dope in the lumen and fiber formation in the spinning duct.

[0079] The main difference between MaSp1 and MaSp2 is the presence of proline (P) residues, which account for 15% of the total amino acid content in MaSp2, while MaSp1 contains no proline. By calculating the number of proline residues in N. clavipes dragline silk, it is possible to estimate the presence of the two proteins in the fiber: 81% MaSp1 and 19% MaSp2. Different spiders have different ratios of MaSp1 and MaSp2. For example, dragline silk fibers from the orb-weaving spider Argiope aurantia contain 41% MaSp1 and 59% MaSp2. Such changes in the ratio of major ampullate gland silk can affect the performance of silk fibers.

[0080] At least seven different types of silk proteins are known for spiders, which weave a single spherical spider web. Silks differ in primary sequence, physical properties, and function. For example, dragline silk, used to construct frames, radii, and lifelines, is known for its remarkable mechanical properties, including strength, toughness, and elasticity. On an equal weight basis, spider silk has a higher toughness than steel and Kevlar®.

[0081] Flagellate silk, found in spiral weft threads, has an extensibility of up to 500%. Minor ampullate silk, found in the auxiliary spirals of spherical spider webs and wrapped around prey, has high tenacity and strength similar to major ampullate silk, but does not hypercontract in water.

[0082] Spider silk is known for its high tensile strength and toughness. Recombinant silk proteins also confer advantageous properties to cosmetic or dermatological compositions, such as improved moisturizing or softening properties, good film-forming properties, and low surface density, among others. The diverse and unique biomechanical properties, along with biocompatibility and a slow degradation rate, make spider silk an excellent candidate for tissue engineering, guided tissue regeneration, and drug delivery biomaterials, cosmetic products (e.g., nail and hair strengtheners, skin care products), and industrial materials (e.g., nanowires, nanofibers, surface coatings).

[0083] In one embodiment, the silk protein may comprise a polypeptide derived from a natural spider silk protein. The polypeptide is not particularly limited as long as it is derived from a natural spider silk protein. Examples of the polypeptide include natural spider silk proteins and recombinant spider silk proteins, such as mutants, analogs, derivatives, or the like of natural spider silk proteins. For superior tenacity, the polypeptide may be derived from a major dragline silk protein produced in the major ampullate gland of spiders. Examples of major ampullate gland spidroins MaSp1 and MaSp2 from Nephila clavipes and ADF3 and ADF4 from Araneus diadematus. Examples of polypeptides derived from major dragline silk proteins include mutants, analogs, derivatives, or the like of major dragline silk proteins. Furthermore, the polypeptide may be derived from a flagellate gland silk protein produced in the flagellate gland of spiders. Examples of flagellate gland silk proteins include flagellate gland silk proteins derived from Nephila clavipes.

[0084] Examples of polypeptides derived from major dragline silk proteins include polypeptides containing two or more units of the amino acid sequence represented by Formula 1:REP1-REP2(1), preferably five or more units thereof, and more preferably ten or more units thereof. Alternatively, a polypeptide derived from a major dragline silk protein may be a polypeptide containing a unit of the amino acid sequence represented by Formula 1:REP1-REP2(1), which has at its C-terminus an amino acid sequence represented by any of SEQ ID NOS: 1-3 of U.S. Patent No. 9,051,453, the entirety of which is incorporated herein by reference, or an amino acid sequence having 90% or greater identity to any of SEQ ID NOS: 1-3 of U.S. Patent No. 9,051,453, the entirety of which is incorporated herein by reference. In polypeptides derived from major dragline silk proteins, the units of the amino acid sequence represented by Formula 1:REP1-REP2(1) may be identical or different from each other. When recombinant proteins are produced using a microorganism such as Escherichia coli as a host, the molecular weight of the polypeptide derived from the major drugline silk protein is, from the viewpoint of productivity, 500 kDa or less, or 300 kDa or less, or 200 kDa or less.

[0085] In formula (1), REP1 represents polyalanine. In REP1, the number of consecutive alanine residues is preferably 2 or more, more preferably 3 or more, even more preferably 4 or more, and particularly preferably 5 or more. Furthermore, in REP1, the number of consecutive alanine residues is preferably 20 or less, more preferably 16 or less, even more preferably 12 or less, and particularly preferably 10 or less. In formula (1), REP2 is an amino acid sequence consisting of 10 to 200 amino acid residues. The total number of glycine, serine, glutamine, and alanine residues contained in the amino acid sequence is 40% or more, preferably 60% or more, and more preferably 70% or more of the total number of amino acid residues contained therein.

[0086] In the primary dragline silk, REP1 corresponds to the crystalline region in the fiber where crystalline β-sheets are formed, while REP2 corresponds to the amorphous region in the fiber where the majority of the components lack regular organization and have more flexibility. Furthermore, [REP1-REP2] corresponds to the repeat region (repeat sequence) composed of crystalline and amorphous regions, which is a characteristic sequence of dragline silk proteins.

[0087] Recombinant silk fragments In some embodiments, recombinant silk protein refers to a recombinant spider silk polypeptide, a recombinant insect silk polypeptide, or a recombinant mussel silk polypeptide. In some embodiments, the recombinant silk protein fragments disclosed herein comprise a recombinant Araneidae or Araneoids spider silk polypeptide or a recombinant Bombyx mori insect silk polypeptide. In some embodiments, the recombinant silk protein fragments disclosed herein comprise a recombinant Araneidae or Araneoids spider silk polypeptide. In some embodiments, the recombinant silk protein fragments disclosed herein comprise a block copolymer having repeating units derived from a natural Araneidae or Araneoids spider silk polypeptide. In some embodiments, the recombinant silk protein fragments disclosed herein comprise a block copolymer having synthetic repeating units derived from an Araneidae or Araneoids spider silk polypeptide and non-repeating units derived from a natural Araneidae or Araneoids spider silk polypeptide.

[0088] Recent advances in genetic engineering have provided a route to producing various types of recombinant silk proteins. Recombinant DNA technology has been used to provide a more practical source of silk proteins. As used herein, "recombinant silk protein" refers to a synthetic protein that is heterologously produced in a prokaryotic or eukaryotic expression system using genetic engineering methods.

[0089] Various methods for synthesizing recombinant silk peptides are known and are described by Ausubel et al., Current Protocols in Molecular Biology § 8 (John Wiley & Sons 1987, (1990)), which is incorporated herein by reference. Escherichia coli, a gram-negative rod-shaped bacterium, is a well-established host for industrial-scale production of proteins. Therefore, the majority of recombinant silk has been produced in E. coli. E. coli is easy to manipulate, has a short generation time, is relatively low cost, and can be scaled up to produce larger amounts of protein.

[0090] Recombinant silk proteins can be produced by transformed prokaryotic or eukaryotic systems containing cDNAs encoding silk proteins, fragments of these proteins, or analogs of such proteins. The recombinant DNA approach allows for the production of recombinant silks with programmed sequences, secondary structures, architectures, and precise molecular weights. There are four major steps in this process: (i) the design and assembly of synthetic silk-like genes into genetic "cassettes," (ii) the insertion of these segments into DNA recombinant vectors, (iii) the transformation of these recombinant DNA molecules into host cells, and (iv) the expression and purification of selected clones.

[0091] The term "recombinant vector," as used herein, includes any vector known to those skilled in the art, including plasmid vectors, cosmid vectors, phage vectors such as lambda phage, viral vectors such as adenovirus or baculovirus vectors, or artificial chromosome vectors such as bacterial artificial chromosomes (BACs), yeast artificial chromosomes (YACs), or P1 artificial chromosomes (PACs). Such vectors include expression vectors and cloning vectors. Expression vectors include plasmids and viral vectors and generally contain a desired coding sequence and appropriate DNA sequences necessary for expression of an operably linked coding sequence in a particular host organism (e.g., bacteria, yeast, or plants) or in an in vitro expression system. Cloning vectors are generally used to manipulate and amplify a specific desired DNA fragment and may lack functional sequences necessary for expression of the desired DNA fragment.

[0092] Prokaryotic systems include gram-negative or gram-positive bacteria. Prokaryotic expression vectors may contain an origin of replication recognizable by the host organism, a homologous or heterologous promoter that is functional in the host, and a DNA sequence encoding a spider silk protein, a fragment of this protein, or a similar protein. Non-limiting examples of prokaryotic expression organisms are cells of Escherichia coli, Bacillus subtilis, Bacillus megaterium, Corynebacterium glutamicum, Anabaena, Caulobacter, Gluconobacter, Rhodobacter, Pseudomonas, Paracoccus, Bacillus (e.g., Bacillus subtilis), Brevibacterium, Corynebacterium, Rhizobium (Sinorhizobium), Flavobacterium, Klebsiella, Enterobacter, Lactobacillus, Lactococcus, Methylobacterium, Propionibacterium, Staphylococcus, or Streptomyces.

[0093] Eukaryotic systems include yeast and insect, mammalian, or plant cells, in which case the expression vector may contain a yeast plasmid origin of replication or an autonomously replicating sequence, a promoter, a DNA sequence encoding the spider silk protein, a fragment, or a similar protein, a polyadenylation sequence, a transcription termination site, and finally, a selection gene. Non-limiting examples of eukaryotic expression organisms include yeast (such as Saccharomyces cerevisiae, Pichia pastoris, basidiosporogenous, ascosporogenous), filamentous fungi (such as Aspergillus niger, Aspergillus oryzae, Aspergillus nidulans, Trichoderma reesei, Acremonium chrysogenum, Candida, Hansenula, Kluyveromyces, Saccharomyces (e.g., Saccharomyces cerevisiae), Schizosaccharomyces, Pichia (e.g., Pichia pastoris), or Yarrowia cells), mammalian cells (such as HeLa cells, COS cells, CHO cells), insect cells (such as Sf9 cells, MEL cells), and "insect host cells" (such as Spodoptera frugiperda or Trichoplusia ni cells). SF9 cells, SF-21 cells, or High-Five cells (SF-9 and SF-21 are ovary cells from Spodoptera frugiperda, and High-Five cells are egg cells from Trichoplusia ni) are "plant host cells" such as tobacco cells, potato cells, or pea cells.

[0094] Various heterologous host systems have been explored to produce different types of recombinant silk. Recombinant partial spidroins as well as engineered silks have been cloned and expressed in bacteria (Escherichia coli), yeast (Pichia pastoris), insects (Bombyx mori larvae), plants (tobacco, soybean, potato, Arabidopsis), mammalian cell lines (BHT / hamster), and transgenic animals (mouse, goat). Most silk proteins are produced with N- or C-terminal His tags to simplify purification and produce sufficient amounts of protein.

[0095] In some embodiments, suitable hosts for expressing recombinant spider silk proteins using heterologous systems may include transgenic animals and plants. In some embodiments, suitable hosts for expressing recombinant spider silk proteins using heterologous systems include bacteria, yeast, and mammalian cell lines. In some embodiments, suitable hosts for expressing recombinant spider silk proteins using heterologous systems include E. coli. In some embodiments, suitable hosts for expressing recombinant spider silk proteins using heterologous systems include transgenic B. mori silkworms generated using genome editing techniques (e.g., CRISPR).

[0096] Recombinant silk proteins in the present disclosure include synthetic proteins based on repeating units of natural silk proteins. In addition to synthetic repetitive silk protein sequences, they can additionally contain one or more naturally occurring non-repetitive silk protein sequences.

[0097] In some embodiments, "recombinant silk protein" refers to recombinant silkworm silk protein or a fragment thereof. Recombinant production of silk fibroin and silk sericin has been reported. Various hosts are used for production, including Escherichia coli, Sacchromyces cerevisiae, Pseudomonas sp., Rhodopseudomonas sp., Bacillus sp., and Strepomyces. See European Patent No. 0230702, incorporated herein by reference in its entirety.

[0098] Also provided herein is the design and biological synthesis of silk fibroin protein-like multiblock polymers containing the GAGAGX (SEQ ID NO: 1) hexapeptide (X is A, Y, V, or S) derived from the repeating domain of the B. mori silk heavy chain (H chain).

[0099] In some embodiments, the present disclosure provides silk protein-like multiblock polymers derived from the repeat domain of B. mori silk heavy chain (H chain) containing GAGAGS (SEQ ID NO: 2) hexapeptide repeat units. The GAGAGS (SEQ ID NO: 2) hexapeptide repeat unit is the core unit of the H chain and plays an important role in the formation of crystalline domains. Silk protein-like multiblock polymers containing GAGAGS (SEQ ID NO: 2) hexapeptide repeat units spontaneously aggregate into a β-sheet structure, similar to natural silk fibroin protein, and have any weight-average molecular weight described herein.

[0100] In some embodiments, the present disclosure provides a silk peptide-like multiblock copolymer composed of a GAGAGS (SEQ ID NO: 2) hexapeptide repeat fragment derived from the heavy chain of a B. mori silk chain and a mammalian elastin VPGVG (SEQ ID NO: 3) motif produced by E. coli. In some embodiments, the present disclosure provides a fusion silk fibroin protein composed of a GAGAGS (SEQ ID NO: 2) hexapeptide repeat fragment derived from the heavy chain of a B. mori silk chain and a GVGVP (SEQ ID NO: 4) motif produced by E. coli, wherein the silk protein-like multiblock copolymer has any weight-average molecular weight described herein.

[0101] In some embodiments, the present disclosure provides (GAGAGS) 16 (SEQ ID NO: 5). In some embodiments, the present disclosure provides a B. mori silkworm recombinant protein composed of a repeat fragment of (GAGAGS) produced by E. coli. 16 (SEQ ID NO: 5) Repeat fragment and non-repetitive (GAGAGS)16 -F-COOH, (GAGAGS) 16 -FF-COOH, (GAGAGS) 16 -FFF-COOH, (GAGAGS) 16 -FFFF-COOH, (GAGAGS) 16 -FFFFFFFF-COOH, (GAGAGS) 16 -FFFFFFFFFFFF-COOH, wherein F has the following amino acid sequence: SGFGPVANGGSGEASSESDFGSSGFGPVANASSGEASSESDFAG (SEQ ID NO: 6), and in a silk protein-like multiblock polymer, has any weight average molecular weight described herein.

[0102] In some embodiments, "recombinant silk protein" refers to a recombinant spider silk protein or a fragment thereof. The production of recombinant spider silk proteins based on partial cDNA clones has been reported. The recombinant spider silk proteins produced as such contain a portion of the repeat sequence derived from the drug-line spider silk protein spidroin 1 from the spider Nephila clavipes. See Xu et al. (Proc. Natl. Acad. Sci. USA, 87:7120-7124 (1990). A cDNA clone encoding a portion of the repeat sequence of spidroin 2, a second fibroin protein from Nephila clavipes dragline silk, and its recombinant synthesis is described in J. Biol. Chem., 1992, volume 267, pp. 19320-19324. Recombinant synthesis of spider silk proteins, including Nephila clavipes protein fragments and variants, from transformed E. coli is described in U.S. Patents 5,728,810 and 5,989,894. A cDNA clone encoding a small ampullate gland spider silk protein and its expression is described in U.S. Patents 5,733,771 and 5,756,677. A cDNA clone encoding a flagellate gland silk protein from a spider that spins a spherical spider web is described in U.S. Patents 5,733,771 and 5,756,677. No. 5,994,099. U.S. Patent No. 6,268,169 describes the recombinant synthesis of spider silk-like proteins derived from repetitive peptide sequences found in the natural spider drug line of Nephila clavipes using E. coli, Bacillus subtilis, and Pichia pastoris recombinant expression systems.WO 03 / 020916 describes cDNA clones encoding spider silk proteins with repetitive sequences derived from the major ampullate glands of Nephila madagascariensis, Nephila senegalensis, Tetragnatha kauaiensis, Tetragnatha versicolor, Argiope aurantia, Argiope trifasciata, Gasteracantha mammosa, and Latrodectus geometricus, the flagellate glands of Argiope trifasciata, the ampullate glands of Dolomedes tenebrosus, two pairs of silk glands from Plectreurys tristis, and the silk glands of the mygalomorph Euagrus chisoseus, and their recombinant production. Each of the above references is incorporated herein by reference in its entirety.

[0103] In some embodiments, the recombinant spider silk protein is a hybrid protein of spider silk protein and insect silk protein, spider silk protein and collagen, spider silk protein and resilin, or spider silk protein and keratin. The spider silk repeating unit comprises or consists of the amino acid sequence of a region comprising or consisting of at least one peptide motif that occurs repeatedly within naturally occurring major ampullate gland polypeptides (such as dragline spider silk polypeptides), minor ampullate gland polypeptides, flagellate gland polypeptides, condensed gland spider silk polypeptides, botryoid gland spider silk polypeptides, or piriform gland spider silk polypeptides.

[0104] In some embodiments, the recombinant spider silk proteins of the present disclosure comprise repeating units of natural spider silk proteins, consensus sequences, and optionally synthetic spider silk proteins derived from one or more natural non-repetitive spider silk protein sequences. The repeating units of natural spider silk polypeptides may comprise dragline spider silk polypeptides or flagellate gland spider silk polypeptides of the Araneidae or Araneoids.

[0105] As used herein, a spider silk "repeating unit" comprises or consists of at least one peptide motif that occurs repeatedly within a naturally occurring major ampullate gland polypeptide (such as a dragline spider silk polypeptide), minor ampullate gland polypeptide, flagellate gland polypeptide, agglutinate gland spider silk polypeptide, botryoid gland spider silk polypeptide, or piriform gland spider silk polypeptide. A "repeating unit" refers to a region of amino acid sequence that corresponds to a region comprising or consisting of at least one peptide motif (e.g., AAAAAA) (SEQ ID NO: 21) or GPGQQ (SEQ ID NO: 16)) that occurs repeatedly within a naturally occurring silk polypeptide (e.g., MaSpI, ADF-3, ADF-4, or Flag) (i.e., the same amino acid sequence), or to an amino acid sequence substantially similar thereto (i.e., a variant amino acid sequence). A "repeat unit" having an amino acid sequence that is "substantially similar" to a corresponding amino acid sequence in a naturally occurring silk polypeptide (i.e., a wild-type repeat unit) is also similar in terms of its properties; for example, a silk protein comprising a "substantially similar repeat unit" is still insoluble and retains its insolubility. For example, a "repeat unit" having an amino acid sequence that is "identical" to the amino acid sequence of a naturally occurring silk polypeptide can be a portion of a silk polypeptide that corresponds to one or more peptide motifs of MaSpI, MaSpII, ADF-3, and / or ADF-4. For example, a "repeat unit" having an amino acid sequence that is "substantially similar" to the amino acid sequence of a naturally occurring silk polypeptide can be a portion of a silk polypeptide that corresponds to one or more peptide motifs of MaSpI, MaSpII, ADF-3, and / or ADF-4, but has one or more amino acid substitutions at specific amino acid positions.

[0106] As used herein, the term "consensus peptide sequence" refers to an amino acid sequence containing a frequently occurring amino acid at a certain position (e.g., G), with other undetermined amino acids being substituted by the placeholder "X." In some embodiments, the consensus sequence is: (i) GPGXX (SEQ ID NO: 7), where X is an amino acid selected from A, S, G, Y, P, and Q; (ii) GGX, where X is an amino acid selected from Y, P, R, S, A, T, N, and Q, preferably Y, P, and Q; (iii) A x , (wherein x is an integer from 5 to 10).

[0107] The consensus peptide sequences GPGXX (SEQ ID NO: 7) and GGX, i.e., glycine-rich motifs, provide flexibility to silk polypeptides and, therefore, to threads formed from silk proteins containing the motifs. Specifically, the repeated GPGXX (SEQ ID NO: 7) motif forms a turned spiral structure, imparting elasticity to silk polypeptides. Both major ampullate and flagellate gland silks contain the GPGXX (SEQ ID NO: 7) motif. The repeated GGX motif is associated with a helical structure with three amino acids per turn and is found in most spider silks. The GGX motif may provide additional elastic properties to silk. The repeated polyalanine Ax (peptide) motif forms a crystalline beta-sheet structure that provides strength to silk polypeptides, as described, for example, in WO 03 / 057727.

[0108] In some embodiments, the recombinant spider silk protein of the present disclosure comprises two identical repeating units, each comprising at least one, and preferably one, amino acid sequence selected from the group consisting of GGRPSDTYG (SEQ ID NO: 8) and GGRPSSSYG (SEQ ID NO: 9) derived from resilin, an elastomeric protein found in most arthropods that provides low stiffness and high strength.

[0109] As used herein, a "non-repetitive unit" refers to an amino acid sequence that is "substantially similar" to the corresponding non-repetitive (carboxy-terminal) amino acid sequence (i.e., the wild-type non-repetitive (carboxy-terminal) unit) within a naturally occurring drug line polypeptide (preferably within the ADF-3, ADF-4, NR3, NR4, ADF-4 of the spider Araneus diadematus, also described in U.S. Pat. No. 9,217,017, incorporated herein by reference in its entirety), and the C16 peptide (spider silk protein eADF4, molecular weight 47.7 kDa, AMSilk) contains 16 repeats of the sequence GSSAAAAAAAASGPGGYGPENQGPSGPGGYGPGGP (SEQ ID NO: 10), an amino acid sequence adapted from the native sequence of ADF4 from A. diadematus. Non-repetitive ADF-4 and its variants display efficient assembly behavior.

[0110] Among the synthetic spider silk proteins, the recombinant silk proteins of the present disclosure in some embodiments comprise the C16 protein having the polypeptide sequence SEQ ID NO: 1, as also described in U.S. Patent No. 8,288,512, which is incorporated herein by reference in its entirety. In addition to the polypeptide sequence set forth in SEQ ID NO: 1, as described in U.S. Patent No. 8,288,512, functional equivalents, functional derivatives, salts, and the like of this sequence, among others, are also included.

[0111] As used herein, "functional equivalents" refers to variants of the above-mentioned amino acid sequences which, at at least one sequence position, have an amino acid other than the amino acid specifically mentioned.

[0112] In some embodiments, the recombinant spider silk proteins of the present disclosure are spidroin major 1 as described by Xu et al., PNAS, USA, 87, 7120, (1990), spidroin major 2 as described by Hinman and Lewis, J. Biol. Chem., 267, 19320, (1922), U.S. Patent Application No. 2016 / 0222174, and U.S. Patent Nos. 9,051,453, 9,617,315, 9,689,089, 8,173,772, 8,642,734, 8,367,803, 8,000,000, and the like. The composition of the present invention comprises an effective amount of at least one natural or recombinant silk protein, including recombinant spider silk proteins such as those described in US Pat. Nos. 97,583, 8,030,024, 7,754,851, 7,148,039, and 7,060,260, or alternatively, spider silk proteins corresponding to the minor spidroins described in patent application WO 95 / 25165. Each of the above-cited references is incorporated herein by reference in its entirety. Additional recombinant spider silk proteins suitable for the recombinant RSPF of the present disclosure include ADF3 and ADF4 from the "major ampullate" gland of Araneus diadematus.

[0113] Recombinant silk is also described in other patents and patent applications, which are incorporated herein by reference: U.S. Patent Publication No. 2004 / 590196, U.S. Patent No. 7,754,851, U.S. Patent Publication No. 2007 / 654470, U.S. Patent No. 7,951,908, U.S. Patent Publication No. 2010 / 785960, U.S. Patent No. 8,034,897, U.S. Patent Publication Nos. 2009 / 0263430, 2008 / 226854, 2009 / 0123967, 2005 / 712095, 2007 / 991037, 2009 / Nos. 0162896, 2008 / 85266, U.S. Patent No. 8,372,436, U.S. Patent Publication Nos. 2007 / 989907, 2009 / 267596, 2010 / 319542, 2009 / 265344, 2012 / 684607, 2004 / 583227, U.S. Patent No. 8,030,024, U.S. Patent Publication No. 2006 / 643569, U.S. Patent No. 7,868,146, U.S. Patent Publication No. 2007 / 991916, U.S. Patent No. 8,097,583, U.S. Patent Publication No. 2006 / 643200 No. 8,729,238, No. 8,877,903, U.S. Patent Publication Nos. 2019 / 0062557, 2016 / 0280960, 2011 / 0201783, 2008 / 991916, 2011 / 986662, 2012 / 697729, 2015 / 0328363, U.S. Patent No. 9,034,816, U.S. Patent Publication No. 2013 / 0172478, U.S. Patent No. 9,217,017, U.S. Patent Publication No. 2017 / 0202995, U.S. Patent No. 8,721,991, U.S. Patent Publication Patent Publication No. 2008 / 227498, U.S. Pat...

Claims

1. A peptide or protein fragment comprising a plurality of amino acids selected from M, R, V, K, T, F, I, L, C, A, Q, Y, N, D, E, G, S, H, P, and W, wherein at least one of the amino acids is modified, substituted, or substituted.

2. 2. The peptide or protein fragment of claim 1, wherein the peptide or protein fragment is a peptide or protein fragment of fibroin, comprising an amino acid modification, substitution, or replacement as defined in claim 1.

3. 3. The peptide or protein fragment of claim 2, wherein the fibroin is a fibroin heavy chain, a fibroin light chain, or a fibrohexamerin.

4. 4. The peptide or protein fragment of claim 1, wherein the peptide or protein fragment comprises from about 2 to about 100 amino acids.

5. 5. The peptide or protein fragment of any one of claims 1 to 4, wherein the peptide or protein fragment comprises 1 to 5 modifications, substitutions and / or alternatives.

6. 6. The peptide or protein fragment of any one of claims 2 to 5, wherein the fibroin is a fibroin heavy chain and the modification, substitution, and / or substitution is at a position corresponding to any one of positions 1 to 5263 of the fibroin heavy chain.

7. 7. The peptide or protein fragment of claim 6, wherein the modification, substitution and / or replacement is selected from the group consisting of an asparagine to aspartic acid modification, substitution and / or replacement, a glutamine to glutamic acid modification, substitution and / or replacement, and a methionine to methionine oxide modification, substitution and / or replacement.

8. 8. The peptide or protein fragment of claim 6 or 7, wherein the modification, substitution and / or substitution is a modification, substitution and / or substitution at Q58, M64, N68, N70, N77, M80, N93, M103, Q125, N132, Q139, Q275, N4191, Q5216 and / or N5262.

9. 6. The peptide or protein fragment of any one of claims 2 to 5, wherein the fibroin is a fibroin light chain and the modification, substitution, and / or substitution is at a position corresponding to any one of positions 1 to 262 of the fibroin light chain.

10. 10. The peptide or protein fragment of claim 9, wherein the modification, substitution and / or replacement is selected from the group consisting of an asparagine to aspartic acid modification, substitution and / or replacement, a glutamine to glutamic acid modification, substitution and / or replacement, and a methionine to methionine oxide modification, substitution and / or replacement.

11. 11. The peptide or protein fragment of claim 9 or 10, wherein the modification, substitution and / or substitution is a modification, substitution and / or substitution at N23, Q24, N28, M69, N105, N108, N118, N136, N138, Q149, N186, N200, Q202, N204, N240, N248 and / or Q255.

12. 6. The peptide or protein fragment of any one of claims 2 to 5, wherein the fibroin is fibrohexamerin (p25) and the modification, substitution, and / or substitution is at a position corresponding to any one of positions 1 to 220 of the fibrohexamerin (p25).

13. 10. The peptide or protein fragment of claim 9, wherein the modification, substitution and / or replacement is selected from the group consisting of an asparagine to aspartic acid modification, substitution and / or replacement, a glutamine to glutamic acid modification, substitution and / or replacement, and a methionine to methionine oxide modification, substitution and / or replacement.

14. 14. A peptide or protein fragment according to claim 12 or 13, wherein the modification, substitution and / or substitution is at Q62, N93, M120, N149, N172, N174 and / or N202.

15. An article comprising a peptide or protein fragment according to any one of claims 1 to 14.

16. 1. A composition comprising a plurality of peptides or protein fragments, each comprising a plurality of amino acids selected from M, R, V, K, T, F, I, L, C, A, Q, Y, N, D, E, G, S, H, P, and W, wherein at least one of the amino acids is modified, substituted, or substituted.

17. 17. The composition of claim 16, wherein the plurality of peptide or protein fragments comprises peptide or protein fragments of fibroin containing amino acid modifications, substitutions, or replacements as defined in claim 16.

18. The composition of claim 17, wherein the fibroin is a fibroin heavy chain, a fibroin light chain, or a fibrohexamerin.

19. The composition of any one of claims 16 to 18, wherein the peptide or protein fragment comprises from about 2 to about 100 amino acids.

20. The composition of any one of claims 1 to 4, wherein the peptide or protein fragment comprises 1 to 5 modifications, substitutions and / or alternatives.

21. The composition of any one of claims 17 to 20, wherein the fibroin is a fibroin heavy chain and the modification, substitution, and / or substitution is at a position corresponding to any one of positions 1 to 5263 of the fibroin heavy chain.

22. 22. The composition of claim 21, wherein the modification, substitution, and / or replacement is selected from the group consisting of an asparagine to aspartic acid modification, substitution, and / or replacement, a glutamine to glutamic acid modification, substitution, and / or replacement, and a methionine to methionine oxide modification, substitution, and / or replacement.

23. 23. The composition of claim 21 or 22, wherein the modification, substitution, and / or substitution is at Q58, M64, N68, N70, N77, M80, N93, M103, Q125, N132, Q139, Q275, N4191, Q5216, and / or N5262.

24. The composition of any one of claims 17 to 20, wherein the fibroin is a fibroin light chain and the modification, substitution, and / or substitution is at a position corresponding to any one of positions 1 to 262 of the fibroin light chain.

25. 25. The composition of claim 24, wherein the modification, substitution, and / or replacement is selected from the group consisting of an asparagine to aspartic acid modification, substitution, and / or replacement, a glutamine to glutamic acid modification, substitution, and / or replacement, and a methionine to methionine oxide modification, substitution, and / or replacement.

26. 26. The composition of claim 24 or 25, wherein the modification, substitution, and / or substitution is at N23, Q24, N28, M69, N105, N108, N118, N136, N138, Q149, N186, N200, Q202, N204, N240, N248, and / or Q255.

27. 21. The composition of any one of claims 17 to 20, wherein the fibroin is a fibrohexamerin (p25) chain and the modification, substitution, and / or substitution is at a position corresponding to any one of positions 1 to 220 of the fibrohexamerin (p25) chain.

28. 28. The composition of claim 27, wherein the modification, substitution, and / or replacement is selected from an asparagine to aspartic acid modification, substitution, and / or replacement, a glutamine to glutamic acid modification, substitution, and / or replacement, and a methionine to methionine oxide modification, substitution, and / or replacement.

29. 29. The composition of claim 27 or 28, wherein the modification, substitution, and / or substitution is at Q62, N93, M120, N149, N172, N174, and / or N202.

30. 30. The composition of any one of claims 17 to 29, wherein each modification, substitution, and / or replacement independently ranges from about 1% to about 99% of the composition.

31. 33. The composition of claim 32, wherein the % modification, substitution, and / or substitution is defined as (the number of peptides or protein fragments containing a modification, substitution, and / or substitution at a particular position divided by the total number of peptides or protein fragments containing said particular position, whether or not they contain a modification, substitution, and / or substitution) x 100.

32. An article comprising the composition of any one of claims 17 to 31.

33. A composition comprising a plurality of peptide or protein fragments of fibroin heavy chain, fibroin light chain, and / or fibrohexamerin (p25), said composition comprising one or more fractions, said plurality of peptide or protein fragments comprising peptide or protein fragments of fibroin containing amino acid modifications, substitutions, or replacements.

34. The plurality of peptides or protein fragments have a weight average molecular weight (M) selected from about 1 kDa to about 20 kDa, about 20 kDa to about 40 kDa, about 40 kDa to about 60 kDa, about 60 kDa to about 80 kDa, about 80 kDa to about 100 kDa, about 100 kDa to about 120 kDa, about 120 kDa to about 140 kDa, or about 140 kDa to about 160 kDa, about 160 kDa to about 180 kDa, about 180 kDa to about 200 kDa, or about 200 kDa to about 250 kDa. ｗ ), and a polydispersity of 1 to about 1.

7.

35. The plurality of peptides or protein fragments have a weight average molecular weight (M) selected from about 10 kDa to about 20 kDa, about 20 kDa to about 40 kDa, about 40 kDa to about 60 kDa, about 60 kDa to about 80 kDa, about 80 kDa to about 100 kDa, about 100 kDa to about 120 kDa, about 120 kDa to about 140 kDa, about 140 kDa to about 160 kDa, or about 160 kDa to about 180 kDa. ｗ ), and a polydispersity of 1 to about 1.1 or 1 to about 1.

2.

36. The plurality of peptides or protein fragments in the fraction have a weight average molecular weight (M) selected from about 10 kDa to about 20 kDa, about 20 kDa to about 40 kDa, about 40 kDa to about 60 kDa, about 60 kDa to about 80 kDa, about 80 kDa to about 100 kDa, about 100 kDa to about 120 kDa, or about 120 kDa to about 140 kDa. ｗ ) and a polydispersity of 1 to about 1.1 or 1 to about 1.

2.

37. The plurality of peptides or protein fragments in the fraction have a weight average molecular weight (M) selected from about 60 kDa to about 80 kDa, about 80 kDa to about 100 kDa, or about 100 kDa to about 120 kDa. ｗ ), and a polydispersity of 1 to about 1.

1.

38. The plurality of peptides or protein fragments in the fraction have a weight average molecular weight (M) selected from about 10 kDa to about 20 kDa, about 20 kDa to about 40 kDa, about 40 kDa to about 60 kDa, about 60 kDa to about 80 kDa, about 80 kDa to about 100 kDa, or about 100 kDa to about 110 kDa. ｗ ) and a polydispersity of 1 to about 1.1 or 1 to about 1.

2.

39. The plurality of peptides or protein fragments in the fraction have a weight average molecular weight (M) selected from about 60 kDa to about 80 kDa, about 80 kDa to about 100 kDa, about 100 kDa to about 120 kDa, or about 120 kDa to about 140 kDa. ｗ ), and a polydispersity of 1 to about 1.

1.

40. The plurality of peptides or protein fragments in the fraction have a weight average molecular weight (M) selected from about 20 kDa to about 40 kDa, or about 40 kDa to about 60 kDa. ｗ ), and a polydispersity of 1 to about 1.1, or 1 to about 1.

2.

41. 34. The composition of claim 33, wherein the one or more fractions are selected from AS77, AS78, AS79, AS80, and AS81.

42. 34. The composition of claim 33, wherein the one or more fractions are selected from AS82, AS83, AS84, AS85, AS86, AS87, AS88, and AS89.

43. 34. The composition of claim 33, wherein the one or more fractions are selected from AS90, AS91, AS92, AS93, and AS94.

44. 34. The composition of claim 33, wherein the one or more fractions are selected from AS95, AS96, AS97, AS98, AS99, and AS100.

45. 34. The composition of claim 33, wherein the plurality of peptides or protein fragments have a weight average molecular weight (Mw) selected from about 40 kDa to about 60 kDa, about 60 kDa to about 80 kDa, about 80 kDa to about 100 kDa, about 100 kDa to about 120 kDa, about 120 kDa to about 140 kDa, about 140 kDa to about 160 kDa, about 160 kDa to about 180 kDa, about 180 kDa to about 200 kDa, or about 200 kDa to about 220 kDa, and a polydispersity of 1 to about 1.

7.

46. The plurality of peptides or protein fragments in the fraction have a weight average molecular weight (M) selected from about 40 kDa to about 60 kDa, about 60 kDa to about 80 kDa, about 80 kDa to about 100 kDa, about 100 kDa to about 120 kDa, about 120 kDa to about 140 kDa, about 140 kDa to about 160 kDa, about 160 kDa to about 180 kDa, about 180 kDa to about 200 kDa, or about 200 kDa to about 210 kDa. ｗ ), and a polydispersity of 1 to about 1.2, or 1 to about 1.

3.

47. The plurality of peptides or protein fragments in the fraction have a weight average molecular weight (M) selected from about 40 kDa to about 60 kDa, about 60 kDa to about 80 kDa, about 80 kDa to about 100 kDa, or about 100 kDa to about 110 kDa, and a polydispersity of 1 to about 1.1, or 1 to about 1.

2. ｗ 34. The composition of claim 33, wherein

48. The plurality of peptides or protein fragments in the fraction have a weight average molecular weight (M) selected from about 60 kDa to about 80 kDa, about 80 kDa to about 100 kDa, about 100 kDa to about 120 kDa, about 120 kDa to about 140 kDa, about 140 kDa to about 160 kDa, about 160 kDa to about 180 kDa, about 180 kDa to about 200 kDa, or about 200 kDa to about 210 kDa. ｗ ), and a polydispersity of 1 to about 1.2, or 1 to about 1.

3.

49. 34. The composition of claim 33, wherein the one or more fractions are selected from AS101, AS102, AS103, AS104, and AS105.

50. 34. The composition of claim 33, wherein the one or more fractions are selected from AS106, AS107, AS108, AS109, AS110, and AS111.

51. 51. The composition of any one of claims 33 to 50, wherein the amino acids are selected from M, R, V, K, T, F, I, L, C, A, Q, Y, N, D, E, G, S, H, P, and W.

52. 52. The composition of any one of claims 33 to 51, wherein the peptide or protein fragment comprises from about 2 to about 100 amino acids.

53. 53. The composition of any one of claims 33 to 52, wherein the peptide or protein fragment comprises 1 to 5 modifications, substitutions and / or alternatives.

54. The composition of any one of claims 33 to 53, wherein the fibroin is a fibroin heavy chain and the modification, substitution, and / or substitution is at a position corresponding to any one of positions 1 to 5263 of the fibroin heavy chain.

55. 54. The composition of any one of claims 33 to 53, wherein the fibroin is a fibroin light chain and the modification, substitution, and / or replacement is at a position corresponding to any one of positions 1 to 262 of the fibroin light chain.

56. 54. The composition of any one of claims 33 to 53, wherein the fibroin is a fibrohexamerin (p25) chain and the modification, substitution, and / or substitution is at a position corresponding to any one of positions 1 to 220 of the fibrohexamerin (p25) chain.

57. 57. The composition of any one of claims 33 to 56, wherein the modification, substitution and / or replacement is selected from: an asparagine to aspartic acid modification, substitution and / or replacement; a glutamine to glutamic acid modification, substitution and / or replacement; and a methionine to methionine oxide modification, substitution and / or replacement.

58. 54. The composition of any one of claims 33 to 53, wherein the modification, substitution, and / or substitution is at a position in the fibroin heavy chain selected from Q58, M64, N68, N70, N77, M80, N93, M103, Q125, N132, Q139, Q275, N4191, Q5216, and / or N5262.

59. 54. The composition of any one of claims 33 to 53, wherein the modification, substitution, and / or substitution is at a fibroin light chain position selected from N23, Q24, N28, M69, N105, N108, N118, N136, N138, Q149, N186, N200, Q202, N204, N240, N248, and / or Q255.

60. 54. The composition of any one of claims 33 to 53, wherein the modification, substitution, and / or substitution is at a fibrohexamerin (p25) position selected from Q62, N93, M120, N149, N172, N174, and / or N202.

61. 61. The composition of any one of claims 33-60, wherein each modification, substitution, and / or replacement independently ranges from about 1% to about 99% of the composition.

62. 62. The composition of any one of claims 33 to 61, wherein the % modification, substitution and / or substitution is defined as (the number of peptides or protein fragments containing a modification, substitution and / or substitution at a particular position divided by the total number of peptides or protein fragments containing said particular position, whether or not they contain a modification, substitution and / or substitution) x 100.

63. 63. The composition of any one of claims 33 to 62, wherein the molecular weight is determined by MALS.

64. An article comprising the composition of any one of claims 33 to 63.

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