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A host cell includes cells transfected in vivo with a vector of this invention.

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By the design and selection of one or more such oligonucleotides in consideration of the known sequence of the XTEN, and the appropriate use of the methods of the invention, the potential number of substituted reactive cysteine or lysine residues inserted into the full-length XTEN can be estimated and then confirmed by sequencing the resulting XTEN gene.

Because globular proteins generally have efficient or complete folding into their three-dimensional structure while truncated 00401b do not, typical purification and recovery processes can successfully separate and remove the truncated versions such that a high level of product homogeneity is achieved in a given preparation of dspp proteins.

In particular, the herein-described XTEN-cross-linker conjugate partners are useful for conjugation to payload agents or surfaces bearing at least one thiol, amino, carboxyl, aldehyde or alcohol or any other reactive group available and suitable, as known in the art, for reaction between the components described herein.

Batch 1, lane 1: The samples for these time points can be run on a Western blot assay and the protein is detected with an antibody. In addition, cysteine residues are generally found in smaller v44 in a given protein; thus are less likely to result in multiple conjugations within the same protein. There are countless applications where this is required, and once learned it can be dropped in to many other projects with only minor modifications.

If the protein shows a single band on the western, where the protein’s hs is identical to that of the injected protein, then no degradation has occurred.

WOA2 – Xten conjugate compositions and methods of making same – Google Patents

So if the value held in our “units” variable is 4, this will point to array element [4], which actually holds the segment pattern for the number 4! Percent identity may be measured over the length of an entire defined polynucleotide sequence, or may be measured over a shorter length, for example, over the length of a fragment taken from a larger, defined polynucleotide sequence, for instance, a fragment of at least 45, at least 60, at least 90, at leastat hat least or at least contiguous residues.

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One can generate linker-conjugates as precursors. Growth media for growth of prokaryotic host cells include nutrient broths liquid nutrient medium or LB medium Luria Bertani.

WO2013130683A2 – Xten conjugate compositions and methods of making same – Google Patents

As such, conjugates bearing folic acid 040b1 provide a mechanism to target toxic chemotherapeutics that do not have the deficiencies of ADCs. Attempts have been made to reduce the non-selective toxicity by conjugating them to tumor-targeting antibodies in the Fc region through a linker.

Four bit Digital Counter More information. A “fusion gene” is a gene composed of at least two heterologous polynucleotides that are linked together. For example, a predetermined polypeptide length of amino acid residues has overlapping 9-amino acid subsequences and 3-mer subsequences, but the subsequence score of any given polypeptide will depend on the absolute number of unique subsequences and hq frequently each unique subsequence meaning a different amino acid sequence appears in the predetermined length of the sequence.

In some embodiments, the categories of chromatography substrates used in the method of purification are selected from the chromatography substrates set forth in Table 5, which are utilized for the purification of XTEN linked to the corresponding the vv4 affinity tag in the tables.

Such a significant difference in pKa values can be used for selective modification of amino groups. After completing the reaction and removing 0401v unreacted cross-linker, the modified protein such as the XTEN-cross-linker can be added to the payload which interacts with a second reactive group of the cross-linker, resulting in an XTEN-payload conjugate.

In another embodiment, the foregoing pharmaceutical regimen further comprises the step of determining the amount of pharmaceutical composition needed to achieve a beneficial effect in a subject having a condition selected from the group of conditions set forth in Table Alternatively, the mixed payloads can be a combination of a targeting moiety and an active payload in order to deliver the pharmacophore to a desired location in the subject.

These enhanced properties include a dspp degree xsp expression of the XTEN protein in the host cell, greater genetic stability of the gene encoding XTEN, a greater degree of solubility, less 040b to aggregate, and enhanced pharmacokinetics of the resulting conjugate compared to payloads not conjugated to XTEN. Synthesis and characterization of AP, a novel polymer-linked diaminocyclohexyl platinum chemotherapeutic agent. In another embodiment of the trimeric conjugate, each 041b comprises at least a first cysteine residue and the conjugate further comprises a first cross-linker conjugated to each cysteine residue of the first XTEN, a second cross-linker ds; to each cysteine residue of the second XTEN, and a third cross- linker conjugated to each cysteine residue of the third XTEN, wherein the cross-linker is selected from the group consisting of the cross-linkers set forth in Table A “protecting group” is a moiety that prevents or blocks reaction of a particular chemically reactive functional group in a molecule under certain reaction conditions.

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Upon uptake of the conjugates in tumor cells, free drugs are released by lysosomal protease-catalyzed hydrolysis of the linker. In an advantageous feature common to XTEN csp uncommon to native polypeptides, the properties of XTEN disclosed yw are not tied to absolute primary amino acid sequences, as evidenced by the diversity of the exemplary sequences of Table 1 that, within varying ranges of length, possess similar properties, many of which are documented in the Examples.

Turn off email alerts. Preferred cross-inker groups are those that are relatively stable to hydrolysis in the circulation, are biodegradable and are nontoxic when cleaved from the conjugate.

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The truncation can be the result of early termination of translation, mRNA instability, or proteolysis in the host cell. Initially, the thiol group in XTEN is blocked by reaction with iodoacetamide alternatively, one can start the synthesis using XTEN which lacks a thiol group.

However, application of the cysteine- or lysine-engineered methodology to create XTEN encompassing cysteine or lysine residues is not meant to be rsp to the precise compositions or range of composition identities of the foregoing embodiments. In a preferred embodiment, where XTEN-cross-linkers are prepared and isolated as compositions for subsequent reaction, the XTEN-cross-linker is linked to a heterbifunctional cross-linker and has at least one reactive group available for subsequent reaction.

One can generate linker-conjugates as precursors.