摘要:
A universal array for multiplexed quantification of variable hemagglutinin such as subcomponents of multivalent annual influenza vaccines that is robust to variations in proteins such as mutations and is capable of quantifying degradation of proteins. Universal capture array (100) comprises one or more substrates (102) and a low-density microarray (104) of sub-arrays (108) comprising spots (106a-c). The microarray (104) is contacted with one or more targets (202) at one or more unknown concentrations, and bound complexes (203) are formed and subsequently quantified with a suitable method. Quantified signals are compared to calibration curves to obtain one or more un-known concentrations and/or quantify degradation of the one or more targets (202). Other embodiments are described and shown.
摘要:
Methods for the quantification of influenza HA proteins and anti-influenza antibodies for the fields of vaccine-related protein quantification, potency determination, and efficacy evaluation are provided. According to the technology, quantification is achieved by providing capture agents attached to an array in a series of decreasing concentrations. Serial dilutions of a reference material also may be introduced. The reference material within each solution binds to the capture agents on the array and is labeled with a label agent capable of producing a detectable signal used to construct a calibration curve. A target material of unknown concentration is introduced to a separate identical array, and the target material binds to the capture agents and also is labeled by a label agent to produce a detectable signal. The calibration curve based on the reference material is then utilized to determine the concentration of the target material without the need to perform replicate experiments.
摘要:
A universal array for multiplexed quantification of variable hemagglutinin such as subcomponents of multivalent annual influenza vaccines that is robust to variations in proteins such as mutations and is capable of quantifying degradation of proteins. Universal capture array ( 100 ) comprises one or more substrates ( 102 ) and a low-density microarray ( 104 ) of sub-arrays ( 108 ) comprising spots ( 106a-c ). The microarray ( 104 ) is contacted with one or more targets ( 202 ) at one or more unknown concentrations, and bound complexes ( 203 ) are formed and subsequently quantified with a suitable method. Quantified signals are compared to calibration curves to obtain one or more unknown concentrations and/or quantify degradation of the one or more targets ( 202 ). Other embodiments are described and shown.
摘要:
Methods for the quantification of influenza HA proteins and anti-influenza antibodies for the fields of vaccine-related protein quantification, potency determination, and efficacy evaluation are provided. According to the technology, quantification is achieved by providing capture agents attached to an array in a series of decreasing concentrations. Serial dilutions of a reference material also may be introduced. The reference material within each solution binds to the capture agents on the array and is labeled with a label agent capable of producing a detectable signal used to construct a calibration curve. A target material of unknown concentration is introduced to a separate identical array, and the target material binds to the capture agents and also is labeled by a label agent to produce a detectable signal. The calibration curve based on the reference material is then utilized to determine the concentration of the target material without the need to perform replicate experiments.
摘要:
Methods for the quantification of influenza HA proteins and anti-influenza antibodies for the fields of vaccine-related protein quantification, potency determination, and efficacy evaluation are provided. According to the technology, quantification is achieved by providing capture agents attached to an array in a series of decreasing concentrations. Serial dilutions of a reference material also may be introduced. The reference material within each solution binds to the capture agents on the array and is labeled with a label agent capable of producing a detectable signal used to construct a calibration curve. A target material of unknown concentration is introduced to a separate identical array, and the target material binds to the capture agents and also is labeled by a label agent to produce a detectable signal. The calibration curve based on the reference material is then utilized to determine the concentration of the target material without the need to perform replicate experiments.
摘要:
There is described a new class or type of initiators for polymerization as a means of signal enhancement, nanoparticle initiators, and methods for amplifying signal resulting from recognition events, thereby enhancing the detection of those recognition events. Methods include amplification achieved through polymerization using a nanoparticle initiator conjugated recognition element that is not consumed during the reaction. The polymer formed as a result of the absorption of light by the nanoparticle initiator and introduction of reactive species into a surrounding polymerizable monomer solution occurs in a spatially-limited region directly surrounding the nanoparticle initiator and is indicative of the recognition event(s). In one embodiment, a semiconductor quantum dot nanoparticle initiator is utilized. In another embodiment, a metal nanoparticle is utilized. In another embodiment, the signal is detected without instrumentation. In yet another embodiment, the signal is detected via a transmission-based instrument which captures an image of the formed polymer.
摘要:
A universal array for multiplexed quantification of variable hemagglutinin such as subcomponents of multivalent annual influenza vaccines that is robust to variations in proteins such as mutations and is capable of quantifying degradation of proteins. Universal capture array (100) comprises one or more substrates (102) and a low-density microarray (104) of sub-arrays (108) comprising spots (106a-c). The microarray (104) is contacted with one or more targets (202) at one or more unknown concentrations, and bound complexes (203) are formed and subsequently quantified with a suitable method. Quantified signals are compared to calibration curves to obtain one or more un-known concentrations and/or quantify degradation of the one or more targets (202). Other embodiments are described and shown.