モノクローナル抗体 作製 受託サービス [ハイブリドーマ]

GL Biochem (Shanghai) Ltd.によるモノクローナル抗体作製の受託サービス[ハイブリドーマ方式]をご案内致します。

ファージディスプレイ方式はこちら。

サービスパッケージの一例

種類	仕様	納品	納期	価格
Monoclonal package 1 (From protein to ascites purification)	SDS-PAGEによる蛋白質チェック 免疫化と血清採取 ELISA同定 融合とhybridoma development アイソタイプ決定 腹水の産生と精製 (2 clones)	2 clones (Petaka) 2-5mgの精製抗体 ELISA結果 アイソタイプ決定結果 プロジェクト報告書	21-25週	こちらをご参照ください。

モノクローナル抗体作製 -基本的な流れ-

Step 1: 抗原の準備

1. タンパク支給の場合: 免疫用で少なくとも5 mg、ELISA用で少なくとも1 mg
   *出来る限り多めに頂けますと成功率UP。
2. ペプチド合成の場合: メーカーで合成 (3-4週) *ご支給も可

Step2: 免疫 (8-10週)

マウス: BALB/C mice, 6-week-oldを6匹

概算日数とイベント:

日数	Process
Day0	50ul pre-immune sera and 1st-immunization with CFA (Complete Freund's adjuvant)
Day21	2nd-immunization with IFA (Incomplete Freund's adjuvant)
Day35	3rd-immunizaiton with IFA
Day45	serum collected for ELISA test
We will provide ELISA data and ELISA protocol to you by e-mail

Maybe more times of immunization are needed for high serum titer. There are 2 weeks' interval between two adjacent immunization.

Step3: ハイブリドーマの作製とスクリーニング (2-3週)

	Process
Day 50	Strengthen-immunization for 2 mice with PBS
Day 54	collection of positive serum and perform two hybridoma fusions based on highest antigen binding titer
Day 64-68	ELISA test for cell culture supernatant and select positive clones; Identify positive antibodies' subtypes
Day 69-83	Provide ELISA data to you for check and we will provide 200ul culture supernatants for each positive hybridoma's. We will keep 10-20 clones of ELISA positive and keep all the positive clones at most of 2 week for message delivery.
Day 84	Make sure the hybridomas the client selected.

Some positive clones will be lost in the process of subcloning and expanding. So we should start 10-15 clones at the beginning for guaranteeing 5-10 clones.

Step4: サブクローニング (4-5週) - 5-10 clones

Day61	1st-subcloning
Day71	ELISA test for cell supernant and select positive
Day72	2nd-subcloning
Day82	ELISA test for cell supernant and select positive
Day83	3rd-subcloning
Day83-day84	ELISA test for cell supernant and select positive
Day84-90	Choose 1 subclone with the highest ELISA titer for each positive hybridoma

Step5: ハイブリドーマの増殖とモノクローナル抗体の産生 (5-10 clones)(4-5週)

Day 91	Injecting 2 mice with liquid paraffin for each clone
Day 91-98	Cells expanding culture
Day99	ELISA test for cell supernant; Identify all the clones still postive
Day92	Injecting the hybridoma to mice intraperitoneally
Day100-106	Collect ascites
Day107-114	Antibody purification by protein A/G and ELISA test

Step6: 同定

ELISA>1:80000(Affinity purified antibody)

5-8 months are needed for all the process in total.




参考価格等はこちら。

関連製品

• ポリクローナル抗体作製
• 抗体作製(ファージディスプレイ)
• シングルドメイン抗体(参考価格)

[お問い合わせ]
メール: biologica [at] biologica.co.jp
電話: 052-262-6732
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