Quantification of residual hydrophobic fusion peptide with monomer and dimer forms using reversed-phase liquid chromatography.
Quantification of residual hydrophobic fusion peptide with monomer and dimer forms using reversed-phase liquid chromatography.
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使用反相液相色谱对单体和二聚体形式的残留疏水性融合肽进行定量。
DOI：
10.1016/j.jchromb.2020.122073
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发表时间：
2020-05-01
期刊：
影响因子：
--
通讯作者：
Paula Lei Q
中科院分区：
文献类型：
--
作者：
Cai CX;Schneck NA;Ivleva VB;Gulla K;Zhang Y;Gowetski D;Paula Lei Q
A fusion peptide mimicking a part of the sequence of HIV-1 envelope glycoprotein with an additional cysteine at its C-terminus (FP8: AVGIGAVFC) was conjugated to a carrier protein through a linker for development of an HIV-1 vaccine. Since this fusion peptide is very hydrophobic with poor solubility and can self-dimerize via a disulfide bond, co-existence of monomeric and dimeric forms presented a major challenge for residual unconjugated FP8 quantification. A reversed-phase liquid chromatography (RPLC) with UV detection was developed to monitor residual FP8 using an experimental correction factor of 0.85 for UV peak area measurement between FP8 dimer and monomer. Therefore, both forms of unconjugated residual FP8 can be measured based on a single FP8 monomer reference curve. Overall, this study demonstrated that the current purification process can remove free residual FP8 to a low level, < 20 μg/mL, which showed negligible impact (< 10%) for the conjugated FP8 ratio measurement using another method, amino acid analysis.
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影响因子：
82.9
作者：
Xu K;Acharya P;Kong R;Cheng C;Chuang GY;Liu K;Louder MK;O'Dell S;Rawi R;Sastry M;Shen CH;Zhang B;Zhou T;Asokan M;Bailer RT;Chambers M;Chen X;Choi CW;Dandey VP;Doria-Rose NA;Druz A;Eng ET;Farney SK;Foulds KE;Geng H;Georgiev IS;Gorman J;Hill KR;Jafari AJ;Kwon YD;Lai YT;Lemmin T;McKee K;Ohr TY;Ou L;Peng D;Rowshan AP;Sheng Z;Todd JP;Tsybovsky Y;Viox EG;Wang Y;Wei H;Yang Y;Zhou AF;Chen R;Yang L;Scorpio DG;McDermott AB;Shapiro L;Carragher B;Potter CS;Mascola JR;Kwong PD
通讯作者：
Kwong PD
DOI：
10.1016/j.jchromb.2019.03.022
发表时间：
2019-05-15
影响因子：
3
作者：
Cai, Cindy X.;Schneck, Nicole A.;Lei, Q. Paula
通讯作者：
Lei, Q. Paula
影响因子：
4.7
作者：
Nahas, Deborah D.;Palladino, Jennifer S.;Hepler, Robert W.
通讯作者：
Hepler, Robert W.
影响因子：
5.6
作者：
Palumbo R;Gogliettino M;Cocca E;Iannitti R;Sandomenico A;Ruvo M;Balestrieri M;Rossi M;Palmieri G
通讯作者：
Palmieri G
DOI：
10.1126/science.aae0474
发表时间：
2016-05-13
期刊：
Science (New York, N.Y.)
影响因子：
--
作者：
Kong R;Xu K;Zhou T;Acharya P;Lemmin T;Liu K;Ozorowski G;Soto C;Taft JD;Bailer RT;Cale EM;Chen L;Choi CW;Chuang GY;Doria-Rose NA;Druz A;Georgiev IS;Gorman J;Huang J;Joyce MG;Louder MK;Ma X;McKee K;O'Dell S;Pancera M;Yang Y;Blanchard SC;Mothes W;Burton DR;Koff WC;Connors M;Ward AB;Kwong PD;Mascola JR
通讯作者：
Mascola JR