Rapid solubility measurement of protein crystals as a function of precipitant concentration with micro-dialysis cell and two-beam interferometer.
A novel dialysis-based technique was developed for the rapid and sample-saving determination of the precipitant dependence of protein solubility. A unique thin dialysis cell was developed which permits both the direct optical measurement of protein concentration and the observation of concentration gradients around a crystal via two-beam interferometry. Interferograms around a lysozyme crystal under the desired concentrations of precipitant (NaCl) and lysozyme were observed to determine whether the crystal is growing or dissolving. This technique enables the precipitant-dependent solubility curve of lysozyme crystal to be obtained in two days using a small sample (total amount: 7 mg lysozyme).
Nakazato
Katsuyoshi
K
Department of Applied Physics, College of Humanities and Sciences, Nihon University, 3-25-40 Sakurajosui, Setagaya-ku, Tokyo, 156-8550, Japan. nakazato@phys.chs.nihon-u.ac.jp
Homma
Tomoko
T
Tomo
Tatsuya
T
D011232:Chemical Precipitation; D003460:Crystallization / Q000379:methods*; D003461:Crystallography / Q000295:instrumentation / Q000379:methods; D019544:Equipment Failure Analysis; D007368:Interferometry / Q000295:instrumentation* / Q000379:methods*; D017551:Microdialysis / Q000295:instrumentation* / Q000379:methods*; D009113:Muramidase / Q000032:analysis / Q000737:chemistry; D011487:Protein Conformation; D011506:Proteins / Q000032:analysis* / Q000737:chemistry*; D012965:Sodium Chloride / Q000032:analysis / Q000737:chemistry; D012995:Solubility; D012996:Solutions; D012997:Solvents / Q000032:analysis / Q000737:chemistry*
D011506:Proteins; D012996:Solutions; D012997:Solvents; D012965:Sodium Chloride; C421864:hen egg lysozyme; D009113:Muramidase