The present disclosure relates to a class of engineered polypeptides and provides a polypeptide comprising the sequence EX.sub.2X.sub.3X.sub.4AX.sub.6X.sub.7EIX.sub.10 X.sub.11LPNLX.sub.16X.sub.17X.sub.18QX.sub.20 X.sub.21AFIX.sub.25X.sub.26LX.sub.28X.sub.29X.sub.30 PX.sub.32QSX.sub.35X.sub.36LLX.sub.39E AKKLX.sub.45X.sub.46X.sub.47Q (SEQ ID NO: 55). The present disclosure also relates to populations of polypeptide variants based on a common scaffold, each polypeptide in the population comprising the amino acid sequence EX.sub.2X.sub.3X.sub.4AX.sub.6X.sub.7EIX.sub.10 X.sub.11LPNLX.sub.16X.sub.17X.sub.18QX.sub.20 X.sub.21AFIX.sub.25X.sub.26LX.sub.28X.sub.29X.sub.30 PX.sub.32QSX.sub.35X.sub.36LLX.sub.39E AKKLX.sub.45X.sub.46X.sub.47Q (SEQ ID NO: 55), and methods for selecting a desired polypeptide having an affinity for a predetermined target from said population.

A Solid Phase Peptide Synthesis (SPPS) device and method of using the same for manufacturing peptides is taught herein. The system comprises at least two reactors, each reactor including a quantity of SPPS resin. The reactors are positioned in series. A de-protecting agent is added to the first reactor and then transferred to the second and third reactors, in series, thereby operating to de-protect the protected N-group. Wash solvent is added to the first reactor and then transferred to the second and this operation repeated several times. Likewise, an amino acid activated ester solution is added, in series, to the first, second and third reactors, thereby operating to couple the amino acid to the de-protected N-group. Wash solvent is added to the first reactor and then transferred to the second and this operation repeated several times prior to the next cycle. The use of the reactors in series reduces the overall solvent required. Online LCMS is also used to monitor progress and identity of reactions happening within the solid phase resin particles.

Systems and methods to characterize dimerization interfaces at the subdomain level of a protein are provided. An exemplary method includes digesting a protein dimer sample into subdomains, labeling the digested protein sample, isolating labeled dimeric and monomeric subdomain fragments, and peptide mapping the labeled sample to determine where the dimer fragments are labeled and where the dimer fragments are not labeled. Regions that show decreased labeling extents in the dimer fraction than that in the monomer fraction are likely involved or in close proximity to the dimerization interface.

Embodiments of the present disclosure pertain to methods of selecting cyclic peptides that bind to a target by transforming a phage display library with a plurality of nucleic acids into bacterial host cells, where the nucleic acids include phage coat protein genes with a combinatorial region that encodes at least one cysteine and at least one non-canonical amino acid. The transformation results in the production of phage particles with phage coat proteins where the cysteine and the non-canonical amino acid couple to one another to form a cyclic peptide library. Phage particles are then screened against the desired target to select bound cyclic peptides. Amino acid sequences of the selected cyclic peptides are then identified. Additional embodiments pertain to methods of constructing a phage display library that encodes the cyclic peptides. Further embodiments of the present disclosure pertain to the produced cyclic peptides, phage display libraries and phage particles.

The invention relates to a peptide combination characterized by peptides each having the same sequence length (SEQL), that can be produced from a mixture (A) comprising a number x of amino acids having a protected acid group or a number z of peptides having an acid group protected by means of a protecting group and an activated amino group, wherein the amino acids are present in the mixture (A) in particular adjustable molar ratios, and a mixture (B) comprising a number y of amino acids having an amino group protected by means of a protecting group, wherein the amino acid molar ratios of the mixture (B) are equal to the amino acid molar ratios of the mixture (A), and wherein the number x=y.

Peptides having activity as protein binding agents are disclosed. The peptides have the following structure (I):

##STR00001##

including stereoisomers, pharmaceutically acceptable salts and prodrugs thereof, wherein R, R.sup.1, L.sup.1, L.sup.2, G, M, Y.sup.1 Y.sup.2 and SEQ are as defined herein. Methods associated with preparation and use of such peptides, as well as pharmaceutical compositions comprising such peptides, are also disclosed.

The invention describes a method for the synthesis of compounds comprising the steps of: (a) compartmentalising two or more sets of primary compounds into microcapsules; such that a proportion of the microcapsules contains two or more compounds; and (b) forming secondary compounds in the microcapsules by chemical reactions between primary compounds from different sets; wherein one or both of steps (a) and (b) is performed under microfluidic control; preferably electronic microfluidic control, The invention further allows for the identification of compounds which bind to a target component of a biochemical system or modulate the activity of the target, and which is co-compartmentalised into the microcapsules.

Disclosed are compositions and method related to variants of SPINK2 that bind to targets other than an endogenous target of SPINK2. In one embodiment, a peptide is provided that comprises the amino acid sequence SEQ ID NO: 1. In further embodiments, an amino acid sequences encoded by nucleotide positions 4 to 42 and/or nucleotide positions 94 to 189 in the nucleotide sequence of SEQ ID NO: 14 flank the amino terminus and the carboxyl terminus, respectively, of the amino acid sequence. In another embodiment, a peptide is provided that comprises an amino acid sequence derived from the amino acid sequence of SEQ ID NO: 1 in which a conservative substitution, deletion, addition and/or insertion of 1 to 5 (inclusive) amino acids has occurred at amino acids other than the 1st Xaa to the 12th Xaa counting from the amino terminus.

The present invention is useful in screening for biomarkers associated with any other disease or condition. Such diseases and conditions range from the neurological diseases, autoimmune diseases and cancers identified above as well as any other disease or condition that has a biomarker such as an antibody or other characterizing protein or biomolecule associated with the disease or progression of the disease. The large ligand libraries of the invention can be used directly in biological fluid, under the appropriate experimental conditions and according to the processes recited herein, to screen for such markers and without the need to use fewer support members (e.g. about 100,000 or less) or without the need to transfer such peptoids or ligands to a microarray before screening the biological fluid. In addition, the ligand libraries may also be used to screen for cell based receptors that specifically relate to a particular cell surface marker.

The invention provides a method of biosynthesis of a cyclic peptide by enzymatically transforming a substrate peptide into the cyclic peptide, wherein the substrate peptide is expressed by a displaying genetic package and comprises at least one Ser or Thr residue and at least one Cys residue, and the enzyme is a post-translationally modifying enzyme (PTME) which is a bifunctional thioether bridge forming dehydratase and cyclase, thereby obtaining the displaying genetic package carrying the cyclic peptide comprising a thioether bridge crosslinking the at least one Ser or Thr to Cys; and further a library of immobilised cyclic peptides, each with a length of at least 10 amino acids, comprising a variety of at least 10.sup.6 library members, which variety comprises at least one of a) a different position of the thioether bridge forming a loop within the substrate peptide; or b) a different number of loops within the substrate peptide.