Reverse transcription-PCR (RT-PCR) was used to amplify either the 550 nt coding for the COOH terminus
of the N gene or the full-length open reading frame for the H gene (14).
Insulin glargine (insulin A21 Gly [right arrow] Asn, and addition of 2 arginine residues to the COOH terminus
of the B chain) had ~22% cross-reactivity with E-test TOSOH II (IRI), whereas it showed concentration-dependent cross-reactivity with the ARCHITECT insulin assay (105% at 10 mIU/L and 83% at 100 mIU/L).
Because different antibodies used in the respective assays show different epitope specificities, they enable detection of either specifically one species, namely A[beta]1-42 [the assay of Innogenetics (11)], or the "family' of A[beta] peptides ending with a COOH terminus
at residue 42 and beginning at different [NH.
RNA was extracted from infected cells by using the guanidinium acid-phenol technique (11), and reverse transcriptase-polymerase chain reaction (RT-PCR) was used to amplify either the 550 nucleotides (nt) coding for the COOH terminus
of N or the full-length open reading frame for H (12).
As is known from investigations on the substrate specificity of tTG (15), this enzyme prefers a Q residue in gliadin for deamidation if it is followed two positions in the direction to the COOH terminus
by a P and then by an F.
1), inferring that there were important antigen determinants at both termini of the N protein but that the antigenicity at the COOH terminus
was higher than that at the N[H.
2] terminus of the BNP peptide, whereas the conjugate antibody is directed at the COOH terminus
To confirm the presence of isoforms of CRT, we performed two-dimensional Western blotting with two different antibodies: monoclonal antibody FMC75, which was produced against recombinant human CRT; and a polyclonal antibody that was produced using synthesized peptides of the human CRT COOH terminus
(amino acids 388-400) as an immunogen.
A third recombinant insulin that is longer acting is insulin glargine (Lantus[R]; Aventis Pharmaceuticals), which differs from regular human insulin by the substitution of glycine for asparagine at position A21 and by the addition of two arginine residues to the COOH terminus
of the B chain.
This problem appeared to be solved with the advent of "intact PTH" two-site IRMAs, which typically use a solid-phase capture antibody directed against the PTH COOH terminus
[usually PTH(39-84)] and a labeled detection antibody raised against PTH(1-34) (11) (see Fig.
The peptides representing the COOH terminus
of the N protein, in particular N371 and N385, had high absorbance/cutoff value ratios with the highest positive detection rate and the lowest hydrophobicity score among all of the synthesized peptides (Fig.
For comparison, we also analyzed the COOH terminus
of the [beta] subunit in that population.