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. 2021 Jul 30;13(8):1503. doi: 10.3390/v13081503

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© 2021 by the authors.

Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).

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Layering of the sucrose gradient and detailed analysis of the gradient: (a) For sucrose gradient ultracentrifugation, the gradient is layered in a SW32Ti tube as displayed; (b) After ultracentrifugation, the first 2 mL fraction from the bottom (fraction 1) is discarded and the next 2 mL fraction (fraction 2) of HBV-rich sucrose is collected; (c) Heparin-affinity chromatography and sucrose gradient ultracentrifugation were performed as described herein. The gradient was fractionated into 2 mL fractions and each fraction was analyzed for protein concentration via Bradford assay, HBsAg via ELISA and HBV GE via qPCR [4,31]. The dotted line indicates the suggested fraction to collect (fraction 2); (d) Similar to panel c, but showing the cumulative percentage of total as well as the percent sucrose concentration (w/v).