Soluble expression in Escherichia coli of functional ScFv of monoclonal antibody MC5 against human colorectal carcinoma
Bibliographic record
Abstract
ObjectiveTo prepare functional soluble single chain va ri able fragment(ScFv) of monoclonal antibody(McAb) MC5 against human colorectal ca rcinoma.Methods mRNA was isolated from the MC5 producing hybr idoma cell line and the DNAs encoding variable domains of heavy and light chains (VH and VL)of the antibody were amplified separately by RT PCR and assembled into ScFv DNA with a linker DNA. The ScFv DNA was ligated into the phagemid vect or pCANTAB5E and the ligated sample was transformed into E.coli TG1. The tra nsfo rmed cells were infected with M13KO7 helper phage to yield recombinant phages. A fter two rounds of panning with human colorectal carci noma cell line SW480 highl y expressing MC5 binding antigen, the phage clones displaying ScFvs of MC5 were selected by ELISA. Two phage clones showing strong signal in ELISA were used to infect E.coli HB2151 to express soluble ScFvs. The soluble ScFvs were ident ified by Dot blot and Western blot, and their antigen binding activities were a ssayed by ELISA.Results VH,VL and ScFv DNAs were about 340, 320 and 750 bp respectively. 10 antigen positive phage clones were selected from 25 preselected phage clones by ELISA. Both the soluble ScFvs derived from the 2 out of the 10 antigen positive phage clones were about 32 000 u and concentra ted in periplasmatic space, and could bind the antigen, and shared the same bind ing site with parental McAb MC5. Conclusion The functional solu ble ScFv of MC5 is successfully produced, which may provide a possible novel tar geting vehicle for in vitro and potential in vivo studies on human color ectal carcinoma.
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How this classification was reachedexpand
Full frame machine prediction
Teacher imitationNot calibrated prevalence, not ground truth. Human validation pending. The Gemma side is a direct model label for every work in the frame, read from the title-only record. The Codex side is a classifier learned from the 10,348 direct Codex labels and calibrated to design-weighted sample rates; fields without enough sample support carry no Codex call. Candidate is the union of the two sides; consensus is their intersection. These outputs are machine_predicted_unvalidated and are not human labels.
Distilled classifier scores by category (both heads)
| Category | Codex | Gemma |
|---|---|---|
| Metaresearch | 0.000 | 0.000 |
| Meta-epidemiology (narrow) | 0.001 | 0.000 |
| Meta-epidemiology (broad) | 0.001 | 0.000 |
| Bibliometrics | 0.000 | 0.001 |
| Science and technology studies | 0.000 | 0.000 |
| Scholarly communication | 0.000 | 0.000 |
| Open science | 0.000 | 0.000 |
| Research integrity | 0.000 | 0.001 |
| Insufficient payload (model declined to judge) | 0.001 | 0.001 |
Machine scores (provisional)
The two teacher heads of the student model, read on this work. A score orders the frame for review; it never asserts a category, and the validation status ships verbatim with every row.
Baseline scores from an immature model (maturity gate not passed, 7 training rounds). Scores rank; they never assert a category.
score_only:v0-immature-baseline · verbatim from the scoring run: score_only means the number may rank works, and no category label ships from itClassification
machine, unvalidatedMachine predicted; a candidate call from one source (direct Gemma or distilled Codex), not a consensus.
How this classification was reached, model by model and score by score, is at the end of the page under "How this classification was reached".