Features

• Animal component-free
• Designed and optimized for small- and large-scale vaccine production
• Scalable, adaptable system to support for Commercialization
• GMP available upon request

Performance

WakoVAC Vero prototypes A and B (media 4 and 5 on the graph) have shown relatively high performances growing larger number of cells under the conditions below.

• Plate	Corning 12 well Plate　(Corning, #3513)
  Cell type	Vero cell
  Number of cells plated	30000 cells/well
  Condition	37℃, 5%CO2
  Culturing duration	5 days
  Media used	2.0 mL

• 

• 1: EMEM/5%FBS 2: DMEM/5%FBS 3: SFM from a competitor 4: WakoVAC Vero prototype A 5: WakoVAC Vero prototype B

Media Optimization Service

WakoVAC Vero will be provided as a made-to-order product, and FUJIFILM Wako is now increasing the variety of prototypes.

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Features

• Animal component-free
• Multiple prototypes are available for testing
• The latest prototype shows an improved virus yield approximately 1.5 times higher than competitor’s Medium A, commercially available serum-free media
• Designed and optimized for small- and large-scale vaccine production
• Compatible with microcarrier
• Adaptation not typically required
• Scalable and adaptable system to support for commercialization
• GMP compliant manufactured

Specification

Intended Use: For Research and Further Manufacturing

Cell culture and target expression

WakoVAC MDCK outperforms FBS contained alternative

Equal to or higher yields of influenza virus than commercially available serum-containing option

Adaptation is not necessarily required

• Switch from a commercially available serum-free medium
  
• Switch from a serum-containing medium
  

Compatible with microcarrier

Media Optimization Service

This is a made-to-order product. Please fill out your information at Media Optimization Service.

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• Trypsin-EDTA Solution without Phenol Red, AF

  for Cell Culture

  Manufacturer : FUJIFILM Wako Pure Chemical Corporation

  Storage Condition : Keep at -20 degrees C.

  Comparison	Product Number	Package Size	Price	Inventory
  	203-20251	100mL	List Price JPY 9,000	Inquire
  	205-20255	500mL	List Price JPY 24,000	In stock in Japan

  CultureSure® Freezing Medium

  for Cell Culture

  Manufacturer : FUJIFILM Wako Pure Chemical Corporation

  Storage Condition : Keep at 2-10 degrees C.

  Comparison	Product Number	Package Size	Price	Inventory
  	Distributor 039-23511	100mL	List Price JPY 8,300	In stock in Japan

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Features

• Support wide range of insect cells
• Performs best with Sf+
  • Cell Growth
  • Cell Viability

   

  

• Fish-derived component(cGMP) and hydrolysate contained
• CoA for non-GMP and GMP available
  Cell growth, cell viability, and protein expression data can be included on CoA upon request
• Intended use: for research and further manufacturing

	Wako Cat# 160-25851	TBD
Format	Liquid	Powder
Package	1L	8kg and/or Customs upon request
Shelf-life	1 year from the date of MFG	2 years from the date of MFG
Storage Condition	2-10˚C, protect from light

Other Information

• Assure lot-to-lot consistency
• Fine protein expression as well with Sf9, Sf21 and Hi5
• L-Glutamine does not need to be supplemented
• Comparative Landscape
  

  Product Description	Supplier	SF	ACF	PF	CD	GMP	Hydrolysate	Cell Type	Shelf-life	Storage	Size	Bag Avaiability
  Sf Insect	FUJIFILM Irvine Scientific	Yes	Yes			Yes	+	Sf9, Sf21	1 year	2-8℃	1L	5L(MTO) Custom upon request
  WakoVAC PSFM-J1	FUJIFILM Wako	Yes	Fish-derived component	Yes		Yes	++	Sf+, Hi5, Sf9, and Sf21	1 year	2-10℃	1L	10L and 20L Powder available

Culturing Sf9 Cells

Retain robust cell growth, viability and target expression with Sf9

Cell Growth	Protein Expression Testing M: Molecular weight size marker Lane 1: Wako VAC PSFM-J1 Lane 2: Medium B Control ←: Target protein The amount of samples being used: 20μl/Lane SDS-PAGE: SuperSep™ Ace 10-20% (Wako Code: 191-15031), 200v constant voltage for 60 minutes Western blotting: PVDF membrane Antibody: Anti- his Tag antibody * Data provided by the Goshima Lab from Nagoya University
Cell Viability

Culturing Hi5 Cells

Retain robust cell growth, viability and target expression with Hi5

Cell Growth

Protein Expression Testing M: Molecular weight size marker Lane 1: Wako VAC PSFM-J1 Lane 2: Wako VAC PSFM-J1 + L-Glutamine Lane 3: Medium C Control + L-Glutamine ←: Target protein The amount of samples being used: 20μl/Lane SDS-PAGE: SuperSep™ Ace 10-20% (Wako Code: 191-15031), 200v constant voltage for 60 minutes Western blotting: PVDF membrane Antibody: Anti- his Tag antibody * Data provided by Mirai-Kougaku

Cell Viability

Target expression data with Sf21

Retain target expression with Sf21

Protein Expression Testing

• The amount of samples being used: 15μl/Lane
• SDS-PAGE: 5-20% gradient gel, 200v constant
• Voltage for 50 minutes

* Data provided by the Goshima Lab from Nagoya University

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Features

• Expressed in E. coli
• Checked residual DNA / Host cell protein
• Missed cleavage rate is lower than competitor‘s Lys-C

Specification

BSA Digestion Analysis

We incubated BSA with each protease for 1 hour and 18 hours.
After incubations, we analized the missed cleavage rate and number of peptides.

➡ The data suggested that our rLys-C have higher activity and specificity in long time reaction than the competitor’s product.

Procedure

1. Sample preparation

①Dissolve or dilute the protein sample to be digested with 25 mmol/L Tris-HCl, 1 mmol/L EDTA，pH 8.5~9.0.
②Add Disulfide threitositol (DTT) or β-mercaptoethanol to the solubilized protein at the final concentration of 5 mmol/L.
③Incubate for 30 min at room temperature.
④Add iodoacetamide to the solubilized protein at the final concentration of 10 mmol/L.
⑤Incubate in dark for 30 min at room temperature.

2. rLys-C preparation

①Dissolve lyophilized powder of rLys-C with 100 μl 12.5 mmol/L Tris-HCl, pH 8.5~9.0.

3. Sample digestion

①Add the prepared rLys-C to sample solution according to the enzyme : protein mass ratio of 1:20~1:100, and incubate at 35℃~37℃ for 2~18 hours.
②The final concentration of 0.5~1.0v/v% trifluoroacetic acid is added to stop the reaction.

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Features

• High specificity and efficiency of protein digestion allow for easy database searches by peptide mass.
• Improved cleavage at lysine residue and increase in the number of peptides are obtained by combination with trypsin.
• Packed in very small quantities according to the amounts used so that sufficient activity for in-gel digestion may be retained.

Application Data

Comparison of In-gel Digestion Using Trypsin (Tp), Lysyl Endopeptidase^® (Lep) and Lep Combined with Tp (Lep +Tp)

BSA band (100ng) resolved by SDS-PAGE was in-gel digested with Tp, Lep and Lep +Tp and analyzed by MALDI-TOFMS. The figure shows the individual mass spectra. The evaluation of these peptidases is summarized in the table.

使用胰蛋白酶 (Tp)、赖氨酰内肽酶^® (Lep) 和 Lep 结合 Tp (Lep +Tp)进行凝胶内消化的比较

通过 SDS-PAGE 解析的 BSA 条带 (100ng) 用 Tp、Lep 和 Lep +Tp 进行凝胶内消化，并通过 MALDI-TOFMS 进行分析。该图显示了单独的质谱图。这些肽酶的评价总结在表中。

Table: Comparison of Trypsin (Tp), Lysyl Endopeptidase^® (Lep) and Lep + Tp

Data provided by: Wada, Y., Osaka Women’s and Children’s Hospital

These results indicate there are very few missed cleavages obtained by Lep digestion. When Tp is used concomitantly with Lep, missed cleavages decrease and the number of identified peptides increase compared to when only Tp is used.

*The value resulted from subtracting the coverage obtained when database searches were performed with Missed cleavage 0 from that obtained when performed with Missed cleavage 1. “Coverage” is the percentage of peptides obtained after in-gel digestion in the whole sequence.

Mass spectrum: Comparison of Trypsin (Tp), Lysyl Endopeptidase^® (Lep) + Tp

References  参考文献

1. Wada, Y. and Kadoya, M.: J. Mass Spectrom., 38, 117(2003).
2. Shevchenko, A., Wilm, M., Vorm, O. and Mann., M.: Anal. Chem., 68, 850(1996).

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beMatrix™ Gelatin

beMatrix™ Gelatin products are created using proprietary technology to yield unmatched safety and quality. Controlled for exceptionally low endotoxin levels and enhanced purity, beMatrix™ products are well suited for a range of biomedical applications including drug excipient, wound dressing materials, bone void fillers, and more.

Features

• Low endotoxin level: NMT 10 EU/g
• Virus inactivated
• IPEC-PQG GMP Compliant
• USP / EP (Ph. Eur.) / JP Compliant
• DMF / MAF registered
• Storage Condition: Room temperature
• Expiration date: 3 years from date of manufacture
• Safety Test performed
  

  Test Item	Cytotoxicity	Sensitization	Intradermal Reaction	Pyrogen	Antigenicity
  Result	Negative	Negative	Negative	Negative	Negative

beMatrix™ Gelatin LS-H

Features

• Alkaline-treated gelatin derived from porcine skin
• High Gel Strength (about 300g)

beMatrix™ Gelatin LS-250

Features

• Sterilized alkaline-treated gelatin derived from porcine skin
• High Gel Strength (about 250g)
• Designed for higher density and faster gelation

beMatrix™ Gelatin LS-W

Features

• Alkaline-treated gelatin derived from porcine skin
• Low Gel Strength (about 100g)

beMatrix™ Gelatin HG

Features

• Sterilized gelatin hydrolyzate derived from porcine skin
• Acute Systemic Toxicity: Negative
• Subacute Toxicity: Negative

beMatrix™ Collagen

The first collagen product used in the world’s first iPS cells based transplant.
beMatrix™ Collagen was used for culturing retinal cells derived from iPS cells that were used for treatment of “age-related macular degeneration”. Cultured cells were successfully transplant in patient without canceration, becoming the world first case.

beMatrix™ Collagen is High quality collagen with strict endotoxin control and virus inactivating processing.

beMatrix™ Collagen AT

Features

• Acid extracted collagen solution derived from porcine tendon
• Low endotoxin level: NMT 0.5 EU/mL
• Storage Condition: Cold storage [4 °C to 8 °C]
• Expiration date: 2 years from the date of manufacture
• Concentration: 3 mg/mL, pH 3

beMatrix™ Collagen TE

Features

• Pepsin-solubilized collagen solution derived from porcine skin
• Low endotoxin level: NMT 0.5 EU/mL
• Storage Condition: Cold storage [4 °C to 8 °C]
• Expiration date: 2 years from the date of manufacture
• Concentration: 5 mg/mL, pH 3

beMatrix™ Collagen FD

Features

• Freeze-dried atelocollagen derived from porcine skin
• Low endotoxin level: NMT 100 EU/g
• Storage Condition: Room temperature
• Expiration date: 2 years from the date of manufacture

Product Appearance

• Spongiform
• Size: approx. 140 x 100 x 10 mm

Application

• Scaffold
• Cell culture / transplantation
• Electro-spinning etc.

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beMatrix Collagen

Low Endotoxin Collagen

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