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The method to research the interaction of drug and ligand
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BSA

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BSA60%80% 66500 Da pKa = 4.6

Ala 48Cys 35Asp 41Glu 58Phe 30Gly 17His 16Ile 15Lys 60Leu 65Met 5Asn 14Pro 28Gln 21Arg 26Ser 32Thr 34Val 38Trp 3Tyr 21

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BSAIII III(AB)BSABSAIIAIIIA

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: BSA CuNiAgAu BSA

1. Ka 2.
3.4.

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270-300nm
5-BSA

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BSA 1*10^-5 M 5 nm 5-HCPT 340 nm 408 nm
2.5 nm 5-HCPTBSABSA

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Non linear chromatography
Frontal Chromatography.

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A new Approach to Determine Camptothecin and Its Analogues Afnity to Human Serum Albumin

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A new Approach to Determine Camptothecin and Its Analogues Afnity to Human Serum Albumin

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Camptothecin (CPT) is mainly from Camptotheca acuminata.A potential treatment for lung cancer, colorectal carcinoma, non-Hodgkins lymphoma, and cervical cancer.
1.Solubility issues.2.Adverse effects.3.Not stable in physiological pH.
PROBLEM
Inactive
Bound to HSA

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Irinotecan and Topotecan treat for metastatic colorectal cancer and refractory ovarian cancer, respectively.
Optical biosensor and spectroscopic techniquesSurface plasmon resonance.Afnity chromatography
Method by
Ka

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1 M Gly
Distribute
PBS

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A1BCA 2 0.16% 0.4 0.95 PH11.25 B4
BCA20-200g/mlBCA0.5-10g/ml
29.5 ug HAS / 2.5 mg MB440 pmol HSA/ 2.5 mg MB

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5 uM ligand 350 ul2.5 mg MB (29.5 ug HSA) 1 ml
Incubated 5 min
Separation1 min
LC-MS-ESI
All of the Rof lactone and carboxylic form of CPT 10-OH-CPT SN-38 AND DB-67 are more than 0.99.
Change the concentration of the ligand:1~ 15 uM

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The free ligandThe blank (MB)
MB

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Camptothecins Short-Term Stability Test:
PBS pH = 7.4 at 18 and 37

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Bmax= 212 15 pmol
1. hydrophobic 2. ionic interaction

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This study conrmed lower binding of inactive carboxylic forms of hydroxycamptothecins analogues to HSA in comparison with CPT. Their lactone analogues could be used as anticancer activity.

2. MB with appropriate immobilized proteins can be used in early stage of durg development.

Additionally, the application of this novel technique as a routine method to accurately qualify interaction between ligand and protein has to be preceded by numerous experiments.

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