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The resin was then collected by centrifugation at 500 g for 10 min and washed in gravity flow column with 5 CVs of buffer containing 20 mM HEPES, pH 7.5, 100 mM NaCl, 10% (v/v) glycerol, 5 mM MgCl 2 , 1 mM MnCl 2 , 25 M TCEP, 0.1% (w/v) LMNG, 0.02% (w/v) CHS and 5 M peptide 20, followed by washing with 15 CVs of buffer containing 20 mM HEPES, pH 7.5, 100 mM NaCl, 10% (v/v) glycerol, 5 mM MgCl 2 , 1 mM MnCl 2 , 25 M TCEP, 0.03% (w/v) LMNG, 0.01% (w/v) GDN, 0.008% (w/v) CHS and 5 M peptide 20

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50 This results in cartilage degradation demonstrable as joint-space narrowing on radiography