1,2-Dichloroethane → Phenobarbital

HRID 1093910

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HRID 1093910 반응식
[Na].O=[C:3]1O[C@H:8]([C@H:10]([CH2:12]O)O)[C:6](O)=[C:4]1O.O[N:15]1[C:19](=[O:20])[CH2:18][CH:17](S(O)(=O)=O)[C:16]1=[O:25].C1C(=O)[N:30](O)[C:28](=[O:29])C1S([O-])(=O)=O.[Na+].[CH2:39](Cl)CCl>O1CCN(CCS(O)(=O)=O)CC1>[CH3:39][CH2:18][C:17]1([C:12]2[CH:10]=[CH:8][CH:6]=[CH:4][CH:3]=2)[C:16](=[O:25])[NH:15][C:19](=[O:20])[NH:30][C:28]1=[O:29]

참가 물질

6

반응물 4

HCID 111,2-DichloroethaneHCID 133909N-HydroxysulfosuccinimideHCID 3520574n-hydroxysulfosuccinimide sodium saltHCID 66586226sodium L-ascorbic acid

생성물 1

HCID 4763Phenobarbital

시약·촉매·용매 1

HCID 781654-Morpholineethanesulfonic acid용매

실험 절차

Ten percent (w/v) latex microparticles (number of carboxy groups approx. 0.21 mmol/g latex, mean microparticle diameter 0.2 μm, Seradyne Inc., Indianapolis, Ind.) were diluted to one percent (w/v) concentration with 10 mM 2-morpholino-ethanesulfonic acid (MES), pH 5.3 containing 0.09% (w/v) ascorbic acid sodium salt. The desired volume of 1% microparticles was measured out and the microparticles activated by the addition of N-hydroxysulfosuccinimide (sulfo-NHS) followed by the addition of N-ethyl-N′-(3-dimethyl-aminopropyl)carbodiimide hydrochloride (EDC.HCl). Both sulfo-NHS and EDC were added at a ratio of 10 moles of each reagent per mole of carboxylates present on the surface of the microparticles. After stirring for about 1 hour at room temperature, the microparticles were washed, concentrated and resuspended to a concentration of 2% by exchange of buffers into 50 mM 3-morpholinopropanesulfonic acid (MOPS), pH 5.7, containing 0.09% (w/v) of ascorbic acid sodium salt, in a hollow-fiber system. A solution of phenobarbital monoclonal antibodies (MAK<Pheba>M-29D4-IgG) in 50 mM MOPS buffer, pH 5.7, also containing 0.09% (w/v) of ascorbic acid sodium salt and 10 g/L of BSA, was added to the resuspended microparticles such that there was 0.2 mg antibody/mL of microparticle solution. The antibody-latex mixture was stirred for about 1 hour at 23-27° C. A solution of 90 mg/mL of BSA in 50 mM MOPS, pH 5.7, containing 0.09% (w/v) of ascorbic acid sodium salt, was then added to the latex mixture (1.125 mg BSA/mg latex). After stirring for 0.5-1.5 hours at 23-27° C., a solution of 11% (w/w) 2-(2-amino-ethoxy)ethanol (AEO) in water adjusted to pH 9 with HCl, was added to the latex mixture. After stirring overnight at 40-45° C., the microparticles were washed with and exchanged into storage buffer (50 mM MOPS, pH 7.4, 0.1% (w/v) BSA, 0.09% sodium azide) to a final concentration of 1% latex microparticles (w/v) which was then stored at 2-8° C. until used.

후처리

  1. washthe microparticles were washed
  2. concentrationconcentrated
  3. additioncontaining 0.09% (w/v) of ascorbic acid sodium salt
  4. additionA solution of phenobarbital monoclonal antibodies (MAK<Pheba>M-29D4-IgG) in 50 mM MOPS buffer, pH 5.7, also containing 0.09% (w/v) of ascorbic acid sodium salt and 10 g/L of BSA
  5. additionwas added to the resuspended microparticles such that there
  6. stirringThe antibody-latex mixture was stirred for about 1 hour at 23-27° C
  7. additionA solution of 90 mg/mL of BSA in 50 mM MOPS, pH 5.7, containing 0.09% (w/v) of ascorbic acid sodium salt
  8. additionwas then added to the latex mixture (1.125 mg BSA/mg latex)
  9. stirringAfter stirring for 0.5-1.5 hours at 23-27° C.
  10. additionwas added to the latex mixture
  11. stirringAfter stirring overnight at 40-45° C.
  12. washthe microparticles were washed with and
  13. customwas then stored at 2-8° C.

출처 및 근거

ORD 기록
ord-5034870a9baa4a109f1f19a89173dee1
출처 데이터셋
uspto-grants-2012_03
특허 번호
US08133743B2

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