The present invention is directed to mutant parvovirus VP1 unique region polypeptides, compositions comprising such polypeptides, methods of making such compositions, as well as methods for identifying the likely presence of parvovirus-neutralizing antibodies, and methods for assessing the functional immunogenicity of parvovirus vaccines and measuring a correlate of efficacy to assess a treatment for parvovirus infection.

A method for measuring cell-mediated immunity to AAV-related gene therapy is provided. The method includes measuring cell-mediated immunity to adeno-associated viruses (AAV) and the transgene carried by these viruses which are being used for gene replacement therapy to treat genetic disorders. These measurements are used to a) estimate the risk of failure, b) the adequacy of immunosuppression, c) adjustment of immunosuppression d) to achieve desired suppression of the immune response, and e) to determine whether a patient about to undergo gene therapy is likely to develop and immune response to the therapy or not. The related embodiment which measures several complement or related proteins on cells is aimed at measuring the risk of TMA, its severity and its response to treatment. The present invention also provides a kit for measuring cell mediated immunity to AAV gene therapy.

Disclosed is a method for detecting a capsid polypeptide of a non-enveloped virus in a sample from a subject which includes (a) contacting the sample with a base, and (b) detecting a capsid polypeptide of the virus in the sample. Moreover, the present disclosure relates to a method for pre-processing a sample from a subject for detection of a virus, including contacting the sample with a base. Moreover, the present disclosure relates to kits, uses and devices related to these methods.

The present technology is directed to methods of preparing affinity chromatographic columns using streptavidin-conjugated particles and biotinylated affinity agents (i.e., biotinylated oligonucleotides, biotinylated antibodies or biotinylated antigen-binding fragments thereof). The methods of the present technology can be utilized to perform affinity capture assays in a high-throughput, efficient manner. Further, the affinity chromatographic columns disclosed herein are customizable due to the ability to utilize any biotinylated affinity agent. The methods herein utilize an on-line loading (i.e., loading the column with biotinylated affinity agent via a liquid chromatography system).

An object of the present invention is to provide a method of identifying an amino acid site in a protein, to which a sugar is bound more accurately than in the related art, and a kit for performing the method. According to the present invention, there is provided a method for identifying an amino acid site in a protein, to which a sugar is bonded, the method including a first mass spectrometry step of subjecting a fragmented protein to mass spectrometry; for a protein in which the bonding of the sugar has been confirmed in the first mass spectrometry step, a substitution-modification step of substituting and/or modifying an amino acid of the protein at the amino acid site in the protein, to which the sugar is bonded and a fragmentation step of fragmenting the protein; and a second mass spectrometry step of subjecting the substituted and/or modified, and fragmented protein obtained by the substitution-modification step and the fragmentation step, to mass spectrometry.

The present invention generally pertains to methods of testing for the presence of anti-drug antibodies (ADAs) against therapeutic viral vectors. In particular, the present invention pertains to the use of biotin-labeled AAV as a capture reagent and ruthenium-labeled AAV as a detection reagent for the detection and quantification of ADAs against AAVs.

The present invention provides methods for determining whether activated carbon can be used for removing viruses or a certain virus from a sample containing a target protein.

The present invention provides methods for identifying, quantifying, and/or characterizing at least one host cell protein (HCP) impurity in a sample containing AAV vectors. The HCP impurities can be enriched through a differential digestion, maintaining intact capsids while exposing the sample to mild denaturation. The mildly denatured sample can subsequently be subjected to enzymatic digestion, generating peptides which can be identified and quantified by liquid chromatography-mass spectrometry (LC-MS) analysis to identify, quantify and/or characterize said at least one HCP impurity.

The present invention is directed to a method of determining DNA concentration in a DNA virus. The invention features three basic steps. The initial step is the specific capture of a defined amount of virus capsid particles on a first solid phase. The second step is lysis of the capsid to release the virus DNA from the first solid phase into a lysis solution. After separating the lysis solution from the first solid phase, the third step is contacting the lysis solution with a second solid phase. The second solid phase captures total DNA derived from the captured capsid. The present invention is also directed to a method for measuring the percentage of full virus capsid, comprising first determining the ssDNA concentration in viruses, and then converting the ssDNA concentration to percentage of full virus capsid using a calibration curve having DNA concentration plotted against standards of % of full capsids.

A method for rapidly detecting viral genome size based on an agarose gel electrophoresis technology belongs to the technical field of virus detection. The kit for detecting the genome size includes a lysis buffer and a sample loading buffer solution, and includes the following raw material components: 5% lauryl sodium sulfate, 0.5 M sodium chloride, 50 mM ethylenediamine tetraacetic acid and 6 DNA gel sample loading buffer dye. By adopting the detection method, a plurality of tedious steps of the existing method are omitted, a clearer gel imaging result can be obtained, the detection time is greatly shortened, and the method is particularly suitable for rapidly detecting virus genome size of parvoviridae family. The method has important application value, and is more in line with the needs of virus detection and quality control research work in the field of gene therapy drug development.