In vitro / In vivo ADME (PK, LC-MS/MS분석)

우정바이오의 ADME 서비스를 통해 약물의 대사 안정성, 약동학적 특성(PK) 등의 확인이 가능하며,
본 서비스는 LC-MS/MS, Microplate reader, and nephelometry를 사용하여 제공하고 있습니다.

주요 서비스

In vitro ADME

Aqueous solubility (Nephelometry)

CYP % Inhibition & IC50 (Microplate reader)

hERG fluorescence polarization assay IC50 (Microplate reader)

Plasma protein binding (LC-MS/MS)

Plasma stability (LC-MS/MS)

Liver microsomal stability (LC-MS/MS)

GI-PAMPA permeability assay (LC-MS/MS)

BBB-PAMPA permeability assay (LC-MS/MS)

Formulation test (Stability, homogeneity)

In vivo ADME (PK: Pharmacokinetics)

Rodent PK study (includes Cassette PK, up to 5 drugs)

Non-rodent PK study: Preliminary investigation in bioequivalence test using

Beagle dog

Formulation test using beagle dog

Dose dependent study

Bioavailability

Gender difference

Food effects

Tissue distribution (includes tumor)

BB assessment(Kp value : Brain to plasma partition coefficient)

Metabolite analysis

Urine, bile, feces analysis

LC-MS/MS 분석

다년간의 동물실험 경험으로 설치류부터 영장류(Global CRO 연계)까지 신속 정확하게 시료 내 (혈액, 조직, 뇨 등) 약물 농도를 측정하는 전문 분석 서비스와 신약개발에 적합한 IonDrive 기술이 탑재된 고성능 LC-MS/MS 장비를 활용하여 천연물 및 대사체,
정량/정성, Method Development와 Validation(MFDS 기준)까지 고객 맞춤형 One stop service를 제공합니다.

주요 장비

• QTRAP 6500+ LC-MS/MS
  [AB SCIEX]
• QTRAP 5500+ LC-MS/MS
  [AB SCIEX]

• IonDrive High Energy Detector+
• Sensitivity (MRM mode)
• Ion source : ESI, APCl

• QTRAP 6500+ LC-MS/MS
  [AB SCIEX]
• QTRAP 5500+ LC-MS/MS
  [AB SCIEX]

• Sensitivity (MRM mode)
• Ion source : ESI, APCl

PK Profile of "Drug A"

핵자기공명(NMR) 분광분석

NMR은 물질의 분자 구조를 분석하는 장비로 신약개발과정에서 꼭 필요하지만 설치 및 유지 관리 측면에서 직접 운용이 어렵습니다. 우정바이오의 약물분석센터를 통해 사용이 가능하며, 직접 사용 서비스도 제공 하고 있어 직접 데이터를 수집할 수 있습니다.

주요 서비스

주요 장비

• AVANCE NEO 400 MHz [BRUKER]

• Magnet field strength : 9.4 Tesla (400 MHz)
• Automation System
  : SampleCase Module
  : Sample diameters from 1 - 10 mm
  : All sample tube length (including Shigemi tubes)
• Resolution for 13 C
  : less than 0.2 Hz at 50 %
  : 2 Hz at 0.55 %, 4 Hz at 0.11 %
• Probehead
  : 5 mm X-nuclei-optimized double resonance broad
  banded probe

STD NMR – binding assay

No.	시료명 (제품명)	Chemical shift (ppm)		Integral[rel]	MoD (%)
1	가교 샘플	BDDE	1.491	0.0437	3.28
		히알루론산	1.890	1.0000

SPR 분석

우정바이오는 2010년부터 쌓아온 수년간의 SPR 분석 노하우를 바탕으로 다양한 물질 특성에 따른 맞춤 상담과 신속 정확한 결과를 제공하고 있습니다. 분석에 사용되는 모든 SPR 시스템은 우정바이오 표준작업지침서에 따라 일일·주간·월간의 장비 유지관리와 함께 SPR 장비 제조회사(Cytiva)의 연 1회 이상 점검을 통해 관리되고 있어 신뢰할 수 있는 데이터 제공이 가능합니다.

주요 서비스

주요 분석물질

• Small molecules
• Biomolecules
  (DNA, RNA, lipids, cells, viruses)
• Transmembrane proteins (TPs)
• Bispecific antibody 
• Immune checkpoint inhibitors (ICIs)
• SARS-CoV-2 Spike  
• Antibody drug conjugates (ADCs) 
• PROTAC 

주요 장비

• Biacore T200 [Cytiva]
• Reichert SR7500D
  [Reichert]

Channel : 4(either parallel or series fluid connection)
Detection: No lower limit for chemical

Sample volume : 2-350 uL
Sample Conc. Range : 1 pM - 1 mM
Sample capacity : Max. 384 (vial or plate)

• Biacore T200 [Cytiva]
• Reichert SR7500D
  [Reichert]

Channel : 2(either parallel or series fluid connection)
Detection : > 100 Da

Sample volume : 1-5000 ul
Sample Conc. Range : 1 pM – 1 mM
Sample capacity : Max. 768 (vial or plate)

SPR(Surface Plasmon Resonance) 광학적 원리를 이용하여 특정 표지 없이 (label-free) 실시간으로 생체분자(Bio-molecules)와 분자 사이의 결합(상호반응)을 검출함으로써 두 분자 간의 affinity(binding parameters(kof, koff, K_D ))를 분석합니다.

Typical SPR sensorgram

Binding Level Screening test

Competitive Inhibition Assay

Protein-protein interactions (PPIs) Inhibitor의 IC50 Values

Fc Receptor Binding Assay

Determination of the Binding of IgGs to FcγR by SPR

Simultaneous Binding of Bispecific Antibody

Reference supported by WOOJUNG BIO

• Front Pharmacol, Allergic Inflammation Caused by Dimerized Translationally Controlled Tumor Protein is Attenuated by Cardamonin. Oct 6, 2021
• Front J Hematol Oncol, KAI1(CD82) is a key molecule to control angiogenesis and switch angiogenic milieu to quiescent state. Sep 16, 2021
• Front Oncol Lett, MED28 increases the colony-forming ability of breast cancer cells by stabilizing the ZNF224 protein upon DNA damage. December 29, 2017
• FrontJ Hematol Oncol, Evaluation of polyphenols from Broussonetia papyrifera as coronavirus protease inhibitors. Jan 22, 2017
• FrontBioorg Med Chem, Dieckol, a SARS-CoV 3CL(pro) inhibitor, isolated from the edible brown algae Ecklonia cava. April 22, 2013
• FrontNature communications, Structural basis for recognition of autophagic receptor NDP52 by the sugar receptor galectin-8. Published 19 Mar, 2013
• FrontNature communications, Protein L-isoaspartyl methyltransferase regulates p53 activity. Published 26 jun, 2012
• FrontAnalytical Biochemistry, Harnessing aptamers for electrochemical detection of endotoxin. 25 February, 2012

보유 장비 리스트

NL-3000 [CYTEK™]

Laser : 405nm, 488nm, 640nm Violet(420 - 829nm), Blue(498 - 829nm), Red(652 - 829nm)  detector module Max color 9 color blue laser panel, 24 colors with three lasers Standard optical configuration Fluorescence linearity FITC R2 ≥ 0.995 / PE R2 ≥ 0.995 Resolution Capable of resolving 0.2 μm beads from noise 12 x 75 mm polystyrene and polypropylene tubes Without  Plate Loader Option Temperature 4 - 30 ℃

EPOCH™2 [BioTEK]

Wavelength range 200 - 999 nm Photometric resolution 0.0001 OD Microplate types 6 - 384 well plates Temperature control 4-Zone incubation to 65 °C Shaking Linear, orbital, double-orbital Detector Photodiode Read methods Endpoint, kinetic, well area scanning

405TS [BioTEK]

Compatible microplates 96 - and 384 - well Shake options 3 speeds Up to 600 sec Precision < 3 % CV @ 300 μL (96-well)≤ 4 % CV @ 80 μL (384-well) Wash volume 25 - 3000 μL

Maximum solvent : 4

System pressure Up to 600 bar Flow range 0.2 to 10 ml/min with G7111A Maximum solve 4 Pump Isocratic, Quaternary, Gradient Well plate MAX 132 Sample Load Detecter Photodiode array

DS-11 [Denovix]

Minimum Sample Size 0.5 μL Detector Type 2048 element CCD Wavelength Range 190 - 840 nm Wavelength Accuracy 0.5 nm Spectral Resolution 1.50 FWHM @253.65 nm Detection Limit 0.04 mg/mL BSA; 0.75 ng/μL dsDNA

YLZ-1248R [Labogene]

Max RPM/RCF Fixed angle 12,000/16,582 xg, Swing out 5,000/5,394 xg Max Capacity Fiixed angle 6*85 ml, Swing out 4*250 ml/250 ml conical Temp range -20 ℃ - 40 ℃ Pulse, timed < 10 hr or continuous

NovoCyte 1000 [ACEA Bioscience]

Laser 488 nm Detector Blue(530/30 nm, 572/28 nm, 675/30 nm) Cell size Blue(530/30 nm, 572/28 nm, 675/30 nm) Fluorescence linearity FITC R2 ≥ 0.995 / PE R2 ≥ 0.995 Resolution Capable of resolving 0.2 μm beads from noise 12 x 75 mm polystyrene and polypropylene tubes Without  Plate Loader Option Temperature 4 - 30 ℃

SpectraMax M2 [Molecular Devices]

Wavelength range 200 - 1000 nm Photometric resolution 0.001 OD Plate formats Cuvette or 6 - 384 well plates Temperature control 4 - 45 °C Shaking 0 to 999 seconds Detector Absorbance(Photomultiplier tube(PMT)) Dual monochromators

NEPHELOsterPlus [BMG Labtech]

Detection modes Nephelometry (light scattering) Measurement modes Endpoint and kinetic Microplate formats 6 - 384 well plates Wavelength Selection Photodiode Wavelength 635±10 nm Incubation +4 °C above ambient up to 45 °C or 60 °C

SpectrumTwo [Perkin Elmer] / MIRacle [PIKE]

Wavelength range 8,300 - 350 cm-1 (KBr), 6,500 - 550 cm-1 (ZnSe) Spectral Resolution 0.5 cm-1 standard S/N Ratio 9,300:1 peak-peak, 5 sec / 32,000:1 peak-peak, 1 min Wavelength Precision 0.1 cm-1 at 3,000 cm-1 Wavelength Accuracy Better than 0.01 cm-1 at 3,000 cm-1 Detector Type DTGS(deuterated triglycine sulfate) ATR Sampling

LightCycler 96 System [Roche]

Range of excitation/emission wavelengths (nm) 470/514 (SYBR, FAM, ResoLight dye) 533/572 (VIC, Hex, Yellow555) 577/620 (Red610, Texas Red), 645/697 (Cy5) Sample format 96-well plates, 8-tube strips Reaction volumes 10 – 50 ul Run time < 40 min for 3-step 40 cycles PCR Programmable temperature range 37 – 98 °C Detecter CCD camera

TurboVap LV [Biotage]

Vial capacity 24 or 48 Sample volume 1.5 - 60 ml Operating Tem 18 - 32 °C Heating Duration 40 min Maximum Pressure 9 bar (0.9 MPa, 130 PSI)