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A list of exemplary cross-linkers is provided in Table 8. The antibody can have one or multiple reactive groups IB. In other embodiments, the invention provides substantially homogeneous polypeptide compositions with XTEN fused to one affinity tag such as, but not limited to the tags of Table 5 by a cleavage sequence linked to either the N- or C-terminus of the XTEN.
WO2013130683A2 – Xten conjugate compositions and methods of making same – Google Patents
It has been difficult to obtain a homogeneous preparation of full-length XTENs due to one or more of the above-mentioned reasons. Phase I clinical and pharmacokinetic study fsp PK1 [N- 2- hydroxypropyl methacrylamide copolymer doxorubicin]: Targeting moieties can be peptides, peptoids, receptor ligands. As such, conjugates bearing folic acid may provide a mechanism to target toxic chemotherapeutics that do not have the deficiencies of ADCs. The invention also contemplates the various combinations of the foregoing conjugates; e.
The “hydrodynamic radius” of a protein affects its rate of diffusion in aqueous solution as well as its ability to migrate in gels of macromolecules.
This enables one to adjust the relative potency or selectivity of the payloads in the resulting XTEN- folate conjugate. Some proteins adopt a random and open, unstructured, or ‘linear’ conformation and as a result have a much larger hydrodynamic radius compared to typical globular proteins of similar molecular weight. Such a uniform distribution of net negative charge in the extended sequence lengths of XTEN also contributes to the unstructured conformation of the polymer that, in turn, rsp result in an effective increase in hydrodynamic radius.
Most proteins have globular structure, which is the most compact three-dimensional structure a protein ddsp have with the smallest hydrodynamic radius. For example, a glycine rich sequence removed from its native coding sequence and operatively linked to a coding sequence other than the native sequence is a heterologous glycine rich sequence. Any of these hww groups can be present or formed on the effector, therapeutic agent, water-soluble polymer, or peptidic carrier.
It has been shown that the acid- sensitive hydrazone bond is the actual cleavage site Jaracz S. These segments are subjected to combinatorial conjugation, enabling the formation of combinatorial products 0401h each library member comprises targeting moieties and payloads. Accordingly, in one embodiment, the invention provides XTEN-cross-linker and XTEN-folate sequences wherein the compositions are conjugated to payloads using a zero-length cross-linkers.
In one embodiment of the XTEN-crosslinker-payload conjugate composition with a first and a second payload, a single second payload is linked to the N-terminus of the XTEN by the second cross-linker conjugated by reaction of an alkyne reactant and an azide reactant selected from the group consisting of the reactants of Table Safety instructions More information.
This conjugate binds to the LHRH -receptor on that is over-expressed on many cancer cells.
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The cyclohexane ring in the spacer arm of this cross-linker decreases the rate of hydrolysis of the maleimide group compared to similar reagents not containing this ring. The foregoing substrates are listed for illustrative purposes and are not intended to limit the scope of substrates that can be employed to practice the invention. A “fusion gene” is a gene composed of at least two heterologous polynucleotides that are linked together. Contaminant components of its natural environment are materials that would typically interfere with diagnostic or therapeutic uses for the polypeptide, and may include enzymes, hormones, and other proteinaceous or non-proteinaceous solutes.
In some embodiments, a selective medium is used which comprises an antibiotic. Conjugations sites of payloads can grouped FIG.
Such properties include but are not limited to secondary or tertiary structure, solubility, protein aggregation, stability, absolute and apparent molecular weight, purity and uniformity, melting properties, contamination and water content. The defined medium may also optionally be supplemented with one or more components from any of the following categories: In one embodiment, XTEN sequences have predominately four to six types of amino acids selected from glycine Galanine Aserine Sthreonine Tglutamate E or proline P that are arranged in a substantially non-repetitive sequence that is about 36 to aboutor about to aboutor about to about amino acid residues in length.
Each library member carries a different ratio of payload A and payload E. In another embodiment of the foregoing XTEN and affinity tag, the composition further comprises one or more helper sequences. Secondary structure elements, such as alpha-helix and beta-sheet, each give rise to a characteristic shape and magnitude of CD spectra, as does the lack of these structure elements. One can generate linker-conjugates as precursors.
In another embodiment, described more fully below, the XTEN incorporates from 1 to about 20, or 1 to about 15, or 1 to about 10, or 1 to 5 cysteine residues wherein the reactive cysteines are utilized for linking to cross-linkers or payloads, as described herein. The most commonly used haloacetyl reagents contain an iodoacetyl group that reacts with sulfhydryl groups at physiological pH.
Once a unique thiol group is introduced in the peptide, it can be selectively modified by compounds containing sufhydryl- reactive such as N-maleimides, haloacetyls, and pyridyl disulfides, as described above. It is intended that a specified sequence mentioned relative to Table 2 has that sequence set forth in Table 2, while a generalized reference to an AE sequence, for example, is intended to encompass any AE sequence having amino acid residues; e.
Methods for identifying agonists of a native polypeptide may comprise contacting a native polypeptide with a candidate agonist molecule and measuring a detectable change in one or more biological activities normally associated with the native polypeptide.