합성 IVT mRNA, circular RNA, self-amplifying RNA 제작서비스

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6.5 - 270 kDa 범위를 커버하는 3가지 컬러 밴드를 포함하며,
최대 100%까지의 transfer 효과를 개런티하는 Prestained protein ladder를 소개합니다.

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IVT mRNA 제작서비스 특징

IVT (in vitro transcribed) mRNA는 cell-free 조건에서 제작되어 세포 주기에 영향을 받지 않으며, 삽입 돌연변이 유발 위험으로부터 안전합니다.
IVT mRNA는 mRNA의 translation 효율과 안정성에 필수적인 5'cap, 3'poly(A) 시퀀스와 같은 mature eukaryotic mRNA 특성을 그대로 재현합니다.
Research grade 뿐 아니라 치료제 모델로 적용할 수 있는 Pre-clinical grade mRNA 제작도 제공해 드립니다.
제공해 주시는 DNA template로 제작하거나, 혹은 DNA 제작부터 의뢰하실 수 있습니다.
일반적인 linear 및 circular, self-amplifying 타입으로 선택하실 수 있습니다.

IVT RNA 종류별 비교

	Linear mRNA	Self-Amplifying RNA	Circular RNA

Structure	Linear; Open 5’ and 3’ end mRNA exonuclease degradation	Linear; Open 5’ and 3’ end mRNA exonuclease degradation	Close loop; No free 5’ 3’ end Eliminating mRNA exonuclease degradation
Ribosome recruiting	5’ Cap: Initiates translation & stabilizes Mrna	5’ Cap: Initiates translation & stabilizes mRNA	nternal Ribosome Entry Site (IRES) : Initiates translation
ORF	• Codon optimization for translation yield • Modified nucleosides, eg. M1Ψ • Insert < 20 kb	• Codon optimization for translation yield • Modified nucleosides, eg. M5C • Insert < 6 kb	• Codon optimization for translation yield • Insert < 4 kb
UTR	Regulatory elements in the 5’ and 3’-UTR stabilize mRNA and increase translation initiation	Replicon for further self-replication of the mRNA	Include IRES + PIE + other elements
PolyA tail	Stabilizes RNA molecule & Prevents degradation	Stabilizes RNA molecule & Prevents degradation	Not required
Protein Expression	+++ Strong	+++++ Strongest	++++ Stronger
Expression half-life	+++ Shorter	+++++ Longest	++++ Longer
Immunogenicity	++++ Low	++ High	+++ Medium
Dosage requirement	++ High	+++++ Lowest	+++ Medium
Novelty	+++ Traditional	++++ Novel	+++++ Novel

Quality Control and Specification

QC	Item	Test Method	Specification	RUO Default	RUO Upgrade	Preclinical Default	Preclinical Upgrade
Identification	Appearance	Visual Inspect	Clear and free of foreign particles	Y	Y	Y	Y
	RNA Length	Capillary Electrophoresis	Target ± 30%	X	Y	Y	Y
	RNA Length	Agarose Gel Electrophoresis	Expected size band detected	Y	X	X	X
	Poly A Length	Enzyme digestion and CE	Target ± 30%	X	Y	Y	Y
	RNA Content	UV Absorbance	Target ± 5%	Y	Y	Y	Y
	PH	pH meter	Target ± 0.5	Y	Y	Y	Y
	Buffer Specification	Client Spec	N/A	Y	Y	Y	Y
Purity	A 260/280 Ratio	UV Spec	1.70 ~ 2.30	Y	Y	Y	Y
	Capping Efficiency	LC-MS	≥ 90%	X	Y	Y	Y
	Size based purity	Capillary Electrophoresis	≥75%	X	Y	Y	Y
Impurity	Total protein residue	Nano Orange Assay	≤ 1%	X	X	Y	Y
	Plasmid DNA Residue	qPCR	≤ 0.1%	X	X	X	Y
	dsRNA	Slot-blot	≤ 1%	X	X	Y	Y
Safety	Endotoxin	Semiquantitative	< 10EU/mg	Y	Y	X	X
	Endotoxin	Quantitative	< 10EU/mg	X	X	Y	Y
	Bioburden	Sdirect Inoculation	No Growth after 48 hrs	X	X	X	Y

mRNA QC 서비스

Option	mRNA QC Test	TAT, days	Amount Required	Method
1	PolyA Tail Length Assay	7	300pmol	RNase digestion and purification to recover the polyA tail fragment, followed by polyA tail length analysis by small RNA bio-analyzer kit assay or LC-MS.
2	Capping Efficiency Assay	14-21	300pmol	RNase digestion to isolate a short oligo from the 5’ end of the mRNA sample to be analyzed by LC-MS to calculate the capping efficiency.
3	dsRNA Residue Assay	10	40ug	Slot-blot assay using a dsRNA-specific antibody to test for dsRNA residues in mRNA sample.
4	RNA Capillary Electrophoresis	5	10ug	Bioanalyzer assay to test RNA length and purity.
5	Total Protein Residue Assay	7	10ug	Nano-orange assay to test the residual protein amount in mRNA sample.
6	Plasmid DNA Residue Assay	14	5ug	q-PCR assay to test the residual plasmid DNA residue amount in mRNA sample.
7	Quantitative Endotoxin Assay	5	5ug	GenScript ToxinSensor™ Chromogenic LAL Endotoxin Assay to test endotoxin level in mRNA sample.
8	BioBurden Assay	14	200ug	TSA plate to recover a broad range of microorganisms such as aerobic and anaerobic bacteria from liquid.

주문정보 - Cat No, PRODUCT, SIZE, 수량 등 항목으로 구성되어있습니다.
	Product	Cat.No.	Size	Maker	Qty	Data Sheet

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합성 IVT mRNA, circular RNA, self-amplifying RNA 제작서비스 | 고마바이오텍