Reduction of Recombinant Adeno-Associated Virus Vector Adsorption on Solid Surfaces by Polyionic Hydrophilic Complex Coating

Abstract

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Surface Modified Nano-Electrospray Needles Improve Sensitivity for Native Mass Spectrometry

“…Both native MS and CD-MS commonly use static nanoelectrospray ionization (nESI) from pulled capillary needles. One possible reason for the limited sensitivity is nonspecific adsorption or other unfavorable interactions with the surface of the borosilicate nESI needles. − A range of important targets are prone to nonspecific adsorption to glass, including plasma proteins, antibodies, lipoproteins, and neuronal peptides. − Due often to their positive charge at neutral pH, these proteins may adsorb and/or unfold on the negatively charged glass surface of nESI needles. ,− Similarly, because AAVs can adsorb to glass, plastic, and metal surfaces, − we hypothesized that the nonspecific adsorption of AAVs with the borosilicate glass needles used for nESI could be limiting the sensitivity of CD-MS.…”

“…These coatings were chosen because they are commercially available, and similar coatings have previously been shown to form inert monolayers that reduce nonspecific adsorption on surfaces. , We then pulled them the coated capillaries into nESI needles (Figure ). Recently, a similar approach used a hydrophilic coating to reduce nonspecific adsorption of AAVs to pipet tubes and quantitative polymerase chain reaction plates . In addition, glass surfaces have previously been passivated for other applications using adsorbed bovine serum albumin (BSA), , PEG, , gold nanoparticles, , and phospholipids .…”

“…Recently, a similar approach used a hydrophilic coating to reduce nonspecific adsorption of AAVs to pipet tubes and quantitative polymerase chain reaction plates. 36 In addition, glass surfaces have previously been passivated for other applications using adsorbed bovine serum albumin (BSA), 32,37 PEG, 23,38 gold nanoparticles, 21,39 and phospholipids. 21 We chose covalent attachment of PEG or PFDCS to the silanol surface to ensure there is no contamination of PEG or PFDCS with the sample.…”

Surface Modified Nano-Electrospray Needles Improve Sensitivity for Native Mass Spectrometry

“…[17][18][19][20] Due often to their positive charge at neutral pH, these proteins may adsorb and/or unfold on the negatively charged glass surface of nESI needles. 14,[16][17][18][19][20][21] Similarly, because AAVs can adsorb to glass, plastic, and metal surfaces, [22][23][24] we hypothesized that the nonspecific adsorption of AAVs with the borosilicate glass needles used for nESI could be limiting the sensitivity of CD-MS.…”

Mass spectrometry friendly pH-gradient anion exchange chromatography for the separation of full and empty adeno-associated virus (AAV) capsids

“…It should be noted that AAVs are prone to non-specific adsorption to solid surfaces which can occur throughout all stages of production and characterization. 28 Materials that were shown to retain AAV particles include plastics, stainless steel and importantly also glass, the material here used to contain AAV samples in the LC autosampler. Variations in peak area potentially result from adsorption to the HPLC vial, rather than being method related, which is also supported by the observed run-to-run decrease in the fluorescence response, as depicted in Fig.…”